anti brca2  (Vector Laboratories)


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    Name:
    Biotinylated Universal Antibody Horse Anti Mouse Rabbit IgG
    Description:
    Biotinylated Universal Antibody Horse Anti Mouse Rabbit IgG is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Universal Horse Anti Mouse Rabbit IgG H L is supplied in liquid format With some exceptions the recommended dilution for most applications is 1 50 H L indicates the antibody recognizes both heavy and light chains This biotinylated Universal secondary antibody is produced in horse and binds equally well to both mouse IgG H L and rabbit IgG H L primary antibodies The high staining quality and sensitivity achieved using this product is equivalent to that obtained with either our biotinylated anti mouse IgG H L or biotinylated anti rabbit IgG H L used individually The convenience of using a single biotinylated secondary antibody for both species of primary and the elimination of possible mistakes in using the wrong secondary antibody is an advantage of this product Note This antibody has significant crossreactivity for rodent IgG and is not recommended for use in staining rodent tissue
    Catalog Number:
    ba-1400
    Price:
    None
    Host:
    Horse
    Size:
    2 1 mg
    Category:
    Antibodies
    Reactivity:
    Universal Mouse Rabbit
    Buy from Supplier


    Structured Review

    Vector Laboratories anti brca2
    Biotinylated Universal Antibody Horse Anti Mouse Rabbit IgG
    Biotinylated Universal Antibody Horse Anti Mouse Rabbit IgG is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Universal Horse Anti Mouse Rabbit IgG H L is supplied in liquid format With some exceptions the recommended dilution for most applications is 1 50 H L indicates the antibody recognizes both heavy and light chains This biotinylated Universal secondary antibody is produced in horse and binds equally well to both mouse IgG H L and rabbit IgG H L primary antibodies The high staining quality and sensitivity achieved using this product is equivalent to that obtained with either our biotinylated anti mouse IgG H L or biotinylated anti rabbit IgG H L used individually The convenience of using a single biotinylated secondary antibody for both species of primary and the elimination of possible mistakes in using the wrong secondary antibody is an advantage of this product Note This antibody has significant crossreactivity for rodent IgG and is not recommended for use in staining rodent tissue
    https://www.bioz.com/result/anti brca2/product/Vector Laboratories
    Average 93 stars, based on 10400 article reviews
    Price from $9.99 to $1999.99
    anti brca2 - by Bioz Stars, 2020-07
    93/100 stars

    Images

    1) Product Images from "Clinical significance of BRCA1 and BRCA2 mRNA and protein expression in patients with sporadic gastric cancer"

    Article Title: Clinical significance of BRCA1 and BRCA2 mRNA and protein expression in patients with sporadic gastric cancer

    Journal: Oncology Letters

    doi: 10.3892/ol.2019.10132

    Immunohistochemistry of BRCA1 and BRCA2 protein expression in gastric cancer tissues. (A) Strong positive BRCA1 staining in the cytoplasm and nuclei, and (B) positive BRCA2 staining in the cytoplasm of gastric adenocarcinoma cells. (C) Negative BRCA1 staining in the cytoplasm and nuclei, and (D) negative BRCA2 staining in the nucleus and cytoplasm of gastric adenocarcinoma cells. (E) Positive BRCA1 staining in the cytoplasm, and (F) positive BRCA2 staining in the cytoplasm of chronic gastritis cells. Magnification, ×400 (scale bar, 50 µm). BRCA1/2, breast cancer type 1 and 2 susceptibility protein.
    Figure Legend Snippet: Immunohistochemistry of BRCA1 and BRCA2 protein expression in gastric cancer tissues. (A) Strong positive BRCA1 staining in the cytoplasm and nuclei, and (B) positive BRCA2 staining in the cytoplasm of gastric adenocarcinoma cells. (C) Negative BRCA1 staining in the cytoplasm and nuclei, and (D) negative BRCA2 staining in the nucleus and cytoplasm of gastric adenocarcinoma cells. (E) Positive BRCA1 staining in the cytoplasm, and (F) positive BRCA2 staining in the cytoplasm of chronic gastritis cells. Magnification, ×400 (scale bar, 50 µm). BRCA1/2, breast cancer type 1 and 2 susceptibility protein.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    Kaplan-Meier survival analysis of patients with gastric cancer. Overall survival according to (A) BRCA1 IHC and (B) BRCA2 IHC. Disease free survival according to (C) BRCA1 IHC and, (D) BRCA2 IHC. Blue line; negative expression of BRCA1/BRCA2 protein by IHC, red line; positive expression of BRCA1/BRCA2 protein by IHC. BRCA1/2, breast cancer type 1 and 2 susceptibility protein; IHC, immunohistochemistry.
    Figure Legend Snippet: Kaplan-Meier survival analysis of patients with gastric cancer. Overall survival according to (A) BRCA1 IHC and (B) BRCA2 IHC. Disease free survival according to (C) BRCA1 IHC and, (D) BRCA2 IHC. Blue line; negative expression of BRCA1/BRCA2 protein by IHC, red line; positive expression of BRCA1/BRCA2 protein by IHC. BRCA1/2, breast cancer type 1 and 2 susceptibility protein; IHC, immunohistochemistry.

    Techniques Used: Immunohistochemistry, Expressing

    2) Product Images from "Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice"

    Article Title: Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4720-05.2006

    D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a biotinylated donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.
    Figure Legend Snippet: D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a biotinylated donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.

    Techniques Used: Immunocytochemistry, Mouse Assay, Knock-Out

    3) Product Images from "gC1qR/p33 Blockade Reduces Staphylococcus aureus Colonization of Target Tissues in an Animal Model of Infective Endocarditis "

    Article Title: gC1qR/p33 Blockade Reduces Staphylococcus aureus Colonization of Target Tissues in an Animal Model of Infective Endocarditis

    Journal:

    doi: 10.1128/IAI.01794-05

    Functional activities of gC1qR MAbs 74.5.2 and 60.11 in rat serum. Biotinylated, clfA -positive bacteria were suspended (10% cell suspension) in selected serum samples from animals treated with gC1qR MAbs. Samples were applied to fibrinogen-coated microtiter
    Figure Legend Snippet: Functional activities of gC1qR MAbs 74.5.2 and 60.11 in rat serum. Biotinylated, clfA -positive bacteria were suspended (10% cell suspension) in selected serum samples from animals treated with gC1qR MAbs. Samples were applied to fibrinogen-coated microtiter

    Techniques Used: Functional Assay

    4) Product Images from "RNase H2 Loss in Murine Astrocytes Results in Cellular Defects Reminiscent of Nucleic Acid-Mediated Autoinflammation"

    Article Title: RNase H2 Loss in Murine Astrocytes Results in Cellular Defects Reminiscent of Nucleic Acid-Mediated Autoinflammation

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00587

    Expression of RNase H2 in the murine central nervous system. (A) Sagittal sections from p21 mouse brains were stained with an RNase H2-specific antiserum followed by a biotinylated secondary antibody, streptavidin-coupled HRP, and developed with the chromogenic substrate 3,3′-Diaminobenzidine (DAB). Note the high expression of RNase H2 in the corpus callosum (cc) and hippocampus, with particularly high expression in the subgranular cell layer (sgcl) of the dentate gyrus (dg). Cornu Ammoni (CA), molecular layer (ml), granular layer (gl). (B) Co-immunofluorescence staining for RNase H2 (red) with the neuronal marker NeuN (green, left panel) and astroglia marker GFAP (green, right panel) shows striking co-localization of RNaseH2 in neuronal cells expressing NeuN in both the cerebral cortex and dg. Scale bar = 150 µm (upper four panels), 10 µm (lower two panels) (C) Co-immunofluorescence staining for RNase H2 shows highest expression of RNase H2 in Ki67-positive proliferating cells. Scale bar = 30 µm.
    Figure Legend Snippet: Expression of RNase H2 in the murine central nervous system. (A) Sagittal sections from p21 mouse brains were stained with an RNase H2-specific antiserum followed by a biotinylated secondary antibody, streptavidin-coupled HRP, and developed with the chromogenic substrate 3,3′-Diaminobenzidine (DAB). Note the high expression of RNase H2 in the corpus callosum (cc) and hippocampus, with particularly high expression in the subgranular cell layer (sgcl) of the dentate gyrus (dg). Cornu Ammoni (CA), molecular layer (ml), granular layer (gl). (B) Co-immunofluorescence staining for RNase H2 (red) with the neuronal marker NeuN (green, left panel) and astroglia marker GFAP (green, right panel) shows striking co-localization of RNaseH2 in neuronal cells expressing NeuN in both the cerebral cortex and dg. Scale bar = 150 µm (upper four panels), 10 µm (lower two panels) (C) Co-immunofluorescence staining for RNase H2 shows highest expression of RNase H2 in Ki67-positive proliferating cells. Scale bar = 30 µm.

    Techniques Used: Expressing, Staining, Immunofluorescence, Marker

    5) Product Images from "Age-related increases in ozone-induced injury and altered pulmonary mechanics in mice with progressive lung inflammation"

    Article Title: Age-related increases in ozone-induced injury and altered pulmonary mechanics in mice with progressive lung inflammation

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00027.2013

    Expression of heme oxygenase-1 (HO-1) in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to HO-1 or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.
    Figure Legend Snippet: Expression of heme oxygenase-1 (HO-1) in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to HO-1 or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.

    Techniques Used: Expressing, Mouse Assay, Staining, Binding Assay

    Expression of mannose receptor in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to mannose receptor or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.
    Figure Legend Snippet: Expression of mannose receptor in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to mannose receptor or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.

    Techniques Used: Expressing, Mouse Assay, Staining, Binding Assay

    Expression of YM-1 in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to YM-1 or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.
    Figure Legend Snippet: Expression of YM-1 in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to YM-1 or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.

    Techniques Used: Expressing, Mouse Assay, Staining, Binding Assay

    Expression of galectin-3 in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to galectin-3 or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.
    Figure Legend Snippet: Expression of galectin-3 in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to galectin-3 or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.

    Techniques Used: Expressing, Mouse Assay, Staining, Binding Assay

    Expression of pro-surfactant protein C (SP-C) in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to pro-SP-C or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Arrowheads indicate insets. Original magnification, ×600; inset, ×1,000.
    Figure Legend Snippet: Expression of pro-surfactant protein C (SP-C) in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to pro-SP-C or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Arrowheads indicate insets. Original magnification, ×600; inset, ×1,000.

    Techniques Used: Expressing, Mouse Assay, Staining, Binding Assay

    Expression of inducible nitric oxide synthase (iNOS) in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to iNOS or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.
    Figure Legend Snippet: Expression of inducible nitric oxide synthase (iNOS) in WT and Sftpd −/− mice. Lung sections, prepared 72 h after exposure of 8-, 27-, and 80-wk-old WT and Sftpd −/− mice to air or ozone, were stained with antibody to iNOS or IgG control followed by biotinylated secondary antibody. Binding was visualized by use of a peroxidase substrate DAB kit. One representative section from 3 separate experiments is shown ( n = 3 mice/treatment group). Original magnification, ×600.

    Techniques Used: Expressing, Mouse Assay, Staining, Binding Assay

    6) Product Images from "Oxidized Phospholipids are Proinflammatory and Proatherogenic in Hypercholesterolemic Mice"

    Article Title: Oxidized Phospholipids are Proinflammatory and Proatherogenic in Hypercholesterolemic Mice

    Journal: Nature

    doi: 10.1038/s41586-018-0198-8

    E06-scFv decreases early aortic valve stenosis, hepatic steatosis, and systemic inflammation a, b, Ldlr −/− (n=11) and Ldlr −/− / E06-scFv (n=10) mice were fed HCD for 15 months and prospectively examined at 3 time points for aortic valve hemodynamics. a , Mean pressure gradients across the aortic valve, determined by Doppler echocardiography. At 12 months there was a 49% lower mean gradient in the Ldlr −/− / E06-scFv mice (2.4±1.9mmHg vs. 4.8±2.4mmHg, mean±SD, p = 0.026, Ldlr −/− / E06-scFv (n=10) and Ldlr −/− (n=9). c, d , Calcification in aortic valve leaflets was determined by von Kossa staining of serial aortic valve sections and AUC compared. AV calcium was reduced in Ldlr −/− / E06-scFv mice by 41.5% (p=0.045, one-tailed-t-test, Ldlr −/− / E06-scFv (n=9) and Ldlr −/− (n=8)). e , Survival of mice used in AV hemodynamic study over 15 months. f , Hepatic cholesterol and triglyceride (TG) levels were reduced by 42% and 47% respectively in Ldlr −/− /E06-scFv mice, Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice g , Livers of mice fed HCD for 16-wks were immunostained with biotinylated E06 IgM (brown) and compared to chow-fed C57BL/6 mice. Shown are representative photomicrographs, representative of 7 Ldlr −/− , 7 Ldlr −/− / E06-scFv and 3 WT (C57BL/6) mice. h , Plasma serum amyloid A (SAA) was decreased 32% in HC fed Ldlr −/− / E06-scFv mice ( Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice).
    Figure Legend Snippet: E06-scFv decreases early aortic valve stenosis, hepatic steatosis, and systemic inflammation a, b, Ldlr −/− (n=11) and Ldlr −/− / E06-scFv (n=10) mice were fed HCD for 15 months and prospectively examined at 3 time points for aortic valve hemodynamics. a , Mean pressure gradients across the aortic valve, determined by Doppler echocardiography. At 12 months there was a 49% lower mean gradient in the Ldlr −/− / E06-scFv mice (2.4±1.9mmHg vs. 4.8±2.4mmHg, mean±SD, p = 0.026, Ldlr −/− / E06-scFv (n=10) and Ldlr −/− (n=9). c, d , Calcification in aortic valve leaflets was determined by von Kossa staining of serial aortic valve sections and AUC compared. AV calcium was reduced in Ldlr −/− / E06-scFv mice by 41.5% (p=0.045, one-tailed-t-test, Ldlr −/− / E06-scFv (n=9) and Ldlr −/− (n=8)). e , Survival of mice used in AV hemodynamic study over 15 months. f , Hepatic cholesterol and triglyceride (TG) levels were reduced by 42% and 47% respectively in Ldlr −/− /E06-scFv mice, Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice g , Livers of mice fed HCD for 16-wks were immunostained with biotinylated E06 IgM (brown) and compared to chow-fed C57BL/6 mice. Shown are representative photomicrographs, representative of 7 Ldlr −/− , 7 Ldlr −/− / E06-scFv and 3 WT (C57BL/6) mice. h , Plasma serum amyloid A (SAA) was decreased 32% in HC fed Ldlr −/− / E06-scFv mice ( Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice).

    Techniques Used: Mouse Assay, Staining, One-tailed Test

    Plasma E06-scFv binds to atherosclerotic lesions and apoptotic thymocytes and is present in aorta of Ldlr −/− /E06-scFv mice a , Staining of atherosclerotic lesions of WHHL rabbit aorta with E06-scFv plasma (left panel), and Ldlr −/− mice (right panel) (both at dilution of 1:20), visualized using biotinylated anti-Myc mAb and ABC-AP VectaStain kit. b , Deconvolution microscopy of E06-scFv plasma (1:20 dilution) binding to apoptotic but not normal cells. Blue, nuclei stained with Hoechst dye; Green, FITC-labeled anti-His tag mAb; Red, Annexin V-PE. c , Binding of E06-scFv plasma (1:20 dilution) to apoptotic thymocytes (7AAD+/Annexin V+) by FACS analysis. d , Expression of E06-scFv in aortic lesion of Ldlr −/− / E06-scFv but not Ldlr −/− mouse. Cross-sections at the AV were stained with biotinylated anti-Myc mAb to identify presence of E06-scFv in atherosclerotic lesion. Nuclei counterstained using Hematoxylin QS (Original ×200). Panels a-c are representative of similar studies with 5 other plasma samples. Panel d is representative of studies in 3 other aortic sections.
    Figure Legend Snippet: Plasma E06-scFv binds to atherosclerotic lesions and apoptotic thymocytes and is present in aorta of Ldlr −/− /E06-scFv mice a , Staining of atherosclerotic lesions of WHHL rabbit aorta with E06-scFv plasma (left panel), and Ldlr −/− mice (right panel) (both at dilution of 1:20), visualized using biotinylated anti-Myc mAb and ABC-AP VectaStain kit. b , Deconvolution microscopy of E06-scFv plasma (1:20 dilution) binding to apoptotic but not normal cells. Blue, nuclei stained with Hoechst dye; Green, FITC-labeled anti-His tag mAb; Red, Annexin V-PE. c , Binding of E06-scFv plasma (1:20 dilution) to apoptotic thymocytes (7AAD+/Annexin V+) by FACS analysis. d , Expression of E06-scFv in aortic lesion of Ldlr −/− / E06-scFv but not Ldlr −/− mouse. Cross-sections at the AV were stained with biotinylated anti-Myc mAb to identify presence of E06-scFv in atherosclerotic lesion. Nuclei counterstained using Hematoxylin QS (Original ×200). Panels a-c are representative of similar studies with 5 other plasma samples. Panel d is representative of studies in 3 other aortic sections.

    Techniques Used: Mouse Assay, Staining, Microscopy, Binding Assay, Labeling, FACS, Expressing

    7) Product Images from "Inflammatory Responses Are Not Sufficient to Cause Delayed Neuronal Death in ATP-Induced Acute Brain Injury"

    Article Title: Inflammatory Responses Are Not Sufficient to Cause Delayed Neuronal Death in ATP-Induced Acute Brain Injury

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0013756

    Death of dopaminergic neurons and microglia in the SNpc induced by ATP. ( A ) ATP (100 nmol in 2 µl PBS) or PBS (2 µl) was unilaterally injected into SNpc (*, injection sites), and brains were obtained after 3 h. Brain sections (30 µm thickness) of the midbrain including the entire SN were prepared, every sixth serial section selected and stained with TH (upper panel) and Iba-1 (lower panel) antibodies, and visualized with biotin-conjugated secondary antibodies and enzymatic detection with the avidin/biotin system unless indicated. At 100 nmol, mild neuronal and microglial damage occurred, thus, 100 nmol ATP was employed for in vivo experiments in this study. Photographs of the most damaged sections were obtained. The contralateral side (contra) and PBS-injected rat brain sections were used as control. ( B ) Serial sections obtained at 3 h were labeled with TH (left panel), Iba-1 (middle panel), and TH/Iba-1 (right panel) antibodies. For visualization of the double-labeling, color reactions using DAB (for TH) and DAB/nickel sulfate (for Iba-1) were applied. Dotted lines indicated damage areas. ( C ) Brain tissue obtained 3 h post ATP treatment was subjected to electron microscopy, as described in “ Materials and Methods ”. Nuclei of neuron (N, white arrow), astrocytes (A, white arrowhead), and microglia (M, black arrow) were shown in intact rat brain whereas cellular structures were severely disrupted in ATP-injected brain. Data in this study are representative of at least 5 animals. Scale bars, 200 µm (A); 100 µm (B); 5 µm (C).
    Figure Legend Snippet: Death of dopaminergic neurons and microglia in the SNpc induced by ATP. ( A ) ATP (100 nmol in 2 µl PBS) or PBS (2 µl) was unilaterally injected into SNpc (*, injection sites), and brains were obtained after 3 h. Brain sections (30 µm thickness) of the midbrain including the entire SN were prepared, every sixth serial section selected and stained with TH (upper panel) and Iba-1 (lower panel) antibodies, and visualized with biotin-conjugated secondary antibodies and enzymatic detection with the avidin/biotin system unless indicated. At 100 nmol, mild neuronal and microglial damage occurred, thus, 100 nmol ATP was employed for in vivo experiments in this study. Photographs of the most damaged sections were obtained. The contralateral side (contra) and PBS-injected rat brain sections were used as control. ( B ) Serial sections obtained at 3 h were labeled with TH (left panel), Iba-1 (middle panel), and TH/Iba-1 (right panel) antibodies. For visualization of the double-labeling, color reactions using DAB (for TH) and DAB/nickel sulfate (for Iba-1) were applied. Dotted lines indicated damage areas. ( C ) Brain tissue obtained 3 h post ATP treatment was subjected to electron microscopy, as described in “ Materials and Methods ”. Nuclei of neuron (N, white arrow), astrocytes (A, white arrowhead), and microglia (M, black arrow) were shown in intact rat brain whereas cellular structures were severely disrupted in ATP-injected brain. Data in this study are representative of at least 5 animals. Scale bars, 200 µm (A); 100 µm (B); 5 µm (C).

    Techniques Used: Injection, Staining, Avidin-Biotin Assay, In Vivo, Labeling, Electron Microscopy

    8) Product Images from "Fermentation with Aquilariae Lignum Enhances the Anti-Diabetic Activity of Green Tea in Type II Diabetic db/db Mouse"

    Article Title: Fermentation with Aquilariae Lignum Enhances the Anti-Diabetic Activity of Green Tea in Type II Diabetic db/db Mouse

    Journal: Nutrients

    doi: 10.3390/nu6093536

    Representative histological images of the insulin-immunoreactive cells in the pancreas, taken from intact normoglycemic or db/db mice. ( A ) Intact control mouse; ( B ) db control mouse; ( C ) metformin 250 mg/kg treated db mouse; ( D ) GT 400 mg/kg treated db mouse; ( E ) fGT 400 mg/kg treated db mouse; ( F ) fGT 200 mg/kg treated db mouse; ( G ) fGT 100 mg/kg treated db mouse. GT, Green tea aqueous lyophilized extracts; fGT, Aquilariae Lignum-fermented green tea aqueous lyophilized extracts; IS, pancreatic islet; PD, pancreatic secretory duct. All immunostained by avidin-biotin-peroxidase complex. Scale bars = 80 µm.
    Figure Legend Snippet: Representative histological images of the insulin-immunoreactive cells in the pancreas, taken from intact normoglycemic or db/db mice. ( A ) Intact control mouse; ( B ) db control mouse; ( C ) metformin 250 mg/kg treated db mouse; ( D ) GT 400 mg/kg treated db mouse; ( E ) fGT 400 mg/kg treated db mouse; ( F ) fGT 200 mg/kg treated db mouse; ( G ) fGT 100 mg/kg treated db mouse. GT, Green tea aqueous lyophilized extracts; fGT, Aquilariae Lignum-fermented green tea aqueous lyophilized extracts; IS, pancreatic islet; PD, pancreatic secretory duct. All immunostained by avidin-biotin-peroxidase complex. Scale bars = 80 µm.

    Techniques Used: Mouse Assay, Avidin-Biotin Assay

    Representative histological images of the glucagon-immunoreactive cells in the pancreas, taken from intact normoglycemic or db/db mice. ( A ) Intact control mouse; ( B ) db control mouse; ( C ) metformin 250 mg/kg treated db mouse; ( D ) GT 400 mg/kg treated db mouse; ( E ) fGT 400 mg/kg treated db mouse; ( F ) fGT 200 mg/kg treated db mouse; ( G ) fGT 100 mg/kg treated db mouse. GT, Green tea aqueous lyophilized extracts; fGT, Aquilariae Lignum-fermented green tea aqueous lyophilized extracts; IS, pancreatic islet; PD, pancreatic secretory duct. All immunostained by avidin-biotin-peroxidase complex. Scale bars = 80 µm.
    Figure Legend Snippet: Representative histological images of the glucagon-immunoreactive cells in the pancreas, taken from intact normoglycemic or db/db mice. ( A ) Intact control mouse; ( B ) db control mouse; ( C ) metformin 250 mg/kg treated db mouse; ( D ) GT 400 mg/kg treated db mouse; ( E ) fGT 400 mg/kg treated db mouse; ( F ) fGT 200 mg/kg treated db mouse; ( G ) fGT 100 mg/kg treated db mouse. GT, Green tea aqueous lyophilized extracts; fGT, Aquilariae Lignum-fermented green tea aqueous lyophilized extracts; IS, pancreatic islet; PD, pancreatic secretory duct. All immunostained by avidin-biotin-peroxidase complex. Scale bars = 80 µm.

    Techniques Used: Mouse Assay, Avidin-Biotin Assay

    9) Product Images from "Improved antifouling properties and selective biofunctionalization of stainless steel by employing heterobifunctional silane-polyethylene glycol overlayers and avidin-biotin technology"

    Article Title: Improved antifouling properties and selective biofunctionalization of stainless steel by employing heterobifunctional silane-polyethylene glycol overlayers and avidin-biotin technology

    Journal: Scientific Reports

    doi: 10.1038/srep29324

    Biofunctionalization of SS-SPB. ( a ) Schematic illustration of the immobilization of bAP on SPB-modified SS via avidin-biotin bridge. ( b ) Spectrophotometric detection of the activity of surface-immobilized bAP. In the negative control, avidin binding sites had been blocked with free biotin to prevent selective attachment. ( c,d ) AFM images showing avidin functionalized SS-SPB. Scale bars 200 nm and 1 μm, respectively. ( e,f ) SS-SPB surface without avidin functionalization. Scale bars as in ( c,d ).
    Figure Legend Snippet: Biofunctionalization of SS-SPB. ( a ) Schematic illustration of the immobilization of bAP on SPB-modified SS via avidin-biotin bridge. ( b ) Spectrophotometric detection of the activity of surface-immobilized bAP. In the negative control, avidin binding sites had been blocked with free biotin to prevent selective attachment. ( c,d ) AFM images showing avidin functionalized SS-SPB. Scale bars 200 nm and 1 μm, respectively. ( e,f ) SS-SPB surface without avidin functionalization. Scale bars as in ( c,d ).

    Techniques Used: Modification, Avidin-Biotin Assay, Activity Assay, Negative Control, Binding Assay

    10) Product Images from "STAT3 inhibition combined with CpG immunostimulation activates antitumor immunity to eradicate genetically distinct castration-resistant prostate cancers"

    Article Title: STAT3 inhibition combined with CpG immunostimulation activates antitumor immunity to eradicate genetically distinct castration-resistant prostate cancers

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-18-1277

    CpG-STAT3ASO conjugate design and cell-selective uptake. (A) Single-stranded CpG-STAT3ASO design; subscript “S” = phosphothioated nucleotides; “o” = C3 units of the carbon linker; red = 2’-O-methyl-modified nucleotides. (B, C) The in vitro uptake of CpG-STAT3ASO Alexa488 compared to STAT3ASO Alexa488 by: (B) primary human immune cells (pDC: CD303 + , mDCs: CD1c + , B cells: CD19 + , and T-cells: CD3 + ); (C) mouse dendritic (DC2.4) and macrophage (RAW264.7) cells, and prostate cancer cells (DU145 and RM9). Cells were incubated for 1 h with 500 nM (B-left panel, C) or with various concentrations (B-right panel) of CpG-STAT3ASO Alexa488 and STAT3ASO Alexa488 without any transfection reagents. Oligonucleotide uptake was measured cytofluorimetrically. (D) CpG-STAT3ASO is internalized by prostate cancer cells via scavenger receptor- and clathrin-dependent endocytosis. DU145 cells were pretreated using various endocytosis inhibitors or placed in 4°C for 1 h before incubation with CpG-STAT3ASO Alexa488 (250 nM) or STAT3ASO Alexa488 (750 nM) for another hour. The percentage of Alexa488-positive cells was assessed by flow cytometry; shown are means+SEM from three independent experiments. (E, F) Partial colocalization of CpG-STAT3ASO with early endosomes and with RNase H1 after cellular uptake. The confocal microscopy to visualize Cy3-labeled oligonucleotides and (E) early endosomal antigen 1 (EEA1) or (F) RNase H1 in prostate cancer cells (DU145) after 15 min and 4 h of incubation with 250 nM CpG-STAT3ASO Cy3 , respectively. (G) The direct interaction of CpG-STAT3ASO Cy3 with RNAse H1 as measured by in situ proximity ligation assay and confocal microscopy. Cells were incubated with 250 nM CpG-STAT3ASO Cy3 or other labeled control oligonucleotides for 4 h before the analysis; shown are representative images from one of three independent experiments.
    Figure Legend Snippet: CpG-STAT3ASO conjugate design and cell-selective uptake. (A) Single-stranded CpG-STAT3ASO design; subscript “S” = phosphothioated nucleotides; “o” = C3 units of the carbon linker; red = 2’-O-methyl-modified nucleotides. (B, C) The in vitro uptake of CpG-STAT3ASO Alexa488 compared to STAT3ASO Alexa488 by: (B) primary human immune cells (pDC: CD303 + , mDCs: CD1c + , B cells: CD19 + , and T-cells: CD3 + ); (C) mouse dendritic (DC2.4) and macrophage (RAW264.7) cells, and prostate cancer cells (DU145 and RM9). Cells were incubated for 1 h with 500 nM (B-left panel, C) or with various concentrations (B-right panel) of CpG-STAT3ASO Alexa488 and STAT3ASO Alexa488 without any transfection reagents. Oligonucleotide uptake was measured cytofluorimetrically. (D) CpG-STAT3ASO is internalized by prostate cancer cells via scavenger receptor- and clathrin-dependent endocytosis. DU145 cells were pretreated using various endocytosis inhibitors or placed in 4°C for 1 h before incubation with CpG-STAT3ASO Alexa488 (250 nM) or STAT3ASO Alexa488 (750 nM) for another hour. The percentage of Alexa488-positive cells was assessed by flow cytometry; shown are means+SEM from three independent experiments. (E, F) Partial colocalization of CpG-STAT3ASO with early endosomes and with RNase H1 after cellular uptake. The confocal microscopy to visualize Cy3-labeled oligonucleotides and (E) early endosomal antigen 1 (EEA1) or (F) RNase H1 in prostate cancer cells (DU145) after 15 min and 4 h of incubation with 250 nM CpG-STAT3ASO Cy3 , respectively. (G) The direct interaction of CpG-STAT3ASO Cy3 with RNAse H1 as measured by in situ proximity ligation assay and confocal microscopy. Cells were incubated with 250 nM CpG-STAT3ASO Cy3 or other labeled control oligonucleotides for 4 h before the analysis; shown are representative images from one of three independent experiments.

    Techniques Used: Modification, In Vitro, Incubation, Transfection, Flow Cytometry, Cytometry, Confocal Microscopy, Labeling, In Situ, Proximity Ligation Assay

    Systemic administration of CpG-STAT3ASO induces regression of bone-localized mouse prostate tumors in immunocompetent mice. C57BL/6 mice were injected intratibially using RM9 or PPS prostate cancer cells. (A) Biodistribution of systemically injected CpG-STAT3ASO Cy3 and STAT3ASO Cy3 in RM9 tumor-bearing mice. Mice were injected IV using 2.5 mg/kg of either oligonucleotide and euthanized 3 h later. Percentages of Cy3 + T-cells (CD3 + ), macrophages (CD11b + F4/80 + ), DCs (CD11b + CD11c + ) and MDSCs (CD11b + /Gr1 + ) were assessed using flow cytometry in single-cell suspensions of bone marrow or spleen. Results of two independent experiments using a total of 6 mice analyzed individually; means+SEM. (B-C) Systemic administration of CpG-STAT3ASO reduces STAT3 activation in bone-localized prostate tumors and in the tumor-associated immune cells. After tumors were established, mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. After the third treatment, mice were euthanized and pSTAT3 activation was assessed in the tumors using immunohistochemistry (B) and flow cytometry (C) in tumor cells (LSC HI SCC HI CD11b - CD3 - ), MDSCs (CD11b + /Gr1 + ), DCs (CD11b + CD11c + ), and T-cells (CD3 + ). C57BL/6 (D-G) or NSG (H) mice were intratibially injected using RM9-Luc or PPS-Luc prostate cancer cells. After tumors were established mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. (D) Tumor progression was monitored using bioluminescent imaging on the AmiX (Spectral Instruments). (E) Repeated systemic administration of CpG-STAT3ASO induces regression of bone-localized tumors and increases the overall survival of mice. Shown are combined results from two independent experiments ( n = 12 mice/each group). (F) Co-injection of CpG ODN and STAT3ASO fails to reproduce the efficacy of the bi-functional CpG-STAT3ASO conjugate against bone-localized RM9-Luc tumors ( n = 6 mice/each group). (G) Systemic administration of CpG-STAT3ASO induced tumor regression in the bone-localized Pten -deficient tumor model (PPS-Luc). Results were combined from two independent experiments ( n = 12 mice/each group). (H) The antitumor effect of CpG-STAT3ASO depended on the presence of an intact immune system and cannot be achieved in immunodeficient NSG mice ( n = 6 mice/each group).
    Figure Legend Snippet: Systemic administration of CpG-STAT3ASO induces regression of bone-localized mouse prostate tumors in immunocompetent mice. C57BL/6 mice were injected intratibially using RM9 or PPS prostate cancer cells. (A) Biodistribution of systemically injected CpG-STAT3ASO Cy3 and STAT3ASO Cy3 in RM9 tumor-bearing mice. Mice were injected IV using 2.5 mg/kg of either oligonucleotide and euthanized 3 h later. Percentages of Cy3 + T-cells (CD3 + ), macrophages (CD11b + F4/80 + ), DCs (CD11b + CD11c + ) and MDSCs (CD11b + /Gr1 + ) were assessed using flow cytometry in single-cell suspensions of bone marrow or spleen. Results of two independent experiments using a total of 6 mice analyzed individually; means+SEM. (B-C) Systemic administration of CpG-STAT3ASO reduces STAT3 activation in bone-localized prostate tumors and in the tumor-associated immune cells. After tumors were established, mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. After the third treatment, mice were euthanized and pSTAT3 activation was assessed in the tumors using immunohistochemistry (B) and flow cytometry (C) in tumor cells (LSC HI SCC HI CD11b - CD3 - ), MDSCs (CD11b + /Gr1 + ), DCs (CD11b + CD11c + ), and T-cells (CD3 + ). C57BL/6 (D-G) or NSG (H) mice were intratibially injected using RM9-Luc or PPS-Luc prostate cancer cells. After tumors were established mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. (D) Tumor progression was monitored using bioluminescent imaging on the AmiX (Spectral Instruments). (E) Repeated systemic administration of CpG-STAT3ASO induces regression of bone-localized tumors and increases the overall survival of mice. Shown are combined results from two independent experiments ( n = 12 mice/each group). (F) Co-injection of CpG ODN and STAT3ASO fails to reproduce the efficacy of the bi-functional CpG-STAT3ASO conjugate against bone-localized RM9-Luc tumors ( n = 6 mice/each group). (G) Systemic administration of CpG-STAT3ASO induced tumor regression in the bone-localized Pten -deficient tumor model (PPS-Luc). Results were combined from two independent experiments ( n = 12 mice/each group). (H) The antitumor effect of CpG-STAT3ASO depended on the presence of an intact immune system and cannot be achieved in immunodeficient NSG mice ( n = 6 mice/each group).

    Techniques Used: Mouse Assay, Injection, Flow Cytometry, Cytometry, Activation Assay, Immunohistochemistry, Imaging, Functional Assay

    11) Product Images from "STAT3 inhibition combined with CpG immunostimulation activates antitumor immunity to eradicate genetically distinct castration-resistant prostate cancers"

    Article Title: STAT3 inhibition combined with CpG immunostimulation activates antitumor immunity to eradicate genetically distinct castration-resistant prostate cancers

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-18-1277

    CpG-STAT3ASO conjugate design and cell-selective uptake. (A) Single-stranded CpG-STAT3ASO design; subscript “S” = phosphothioated nucleotides; “o” = C3 units of the carbon linker; red = 2’-O-methyl-modified nucleotides. (B, C) The in vitro uptake of CpG-STAT3ASO Alexa488 compared to STAT3ASO Alexa488 by: (B) primary human immune cells (pDC: CD303 + , mDCs: CD1c + , B cells: CD19 + , and T-cells: CD3 + ); (C) mouse dendritic (DC2.4) and macrophage (RAW264.7) cells, and prostate cancer cells (DU145 and RM9). Cells were incubated for 1 h with 500 nM (B-left panel, C) or with various concentrations (B-right panel) of CpG-STAT3ASO Alexa488 and STAT3ASO Alexa488 without any transfection reagents. Oligonucleotide uptake was measured cytofluorimetrically. (D) CpG-STAT3ASO is internalized by prostate cancer cells via scavenger receptor- and clathrin-dependent endocytosis. DU145 cells were pretreated using various endocytosis inhibitors or placed in 4°C for 1 h before incubation with CpG-STAT3ASO Alexa488 (250 nM) or STAT3ASO Alexa488 (750 nM) for another hour. The percentage of Alexa488-positive cells was assessed by flow cytometry; shown are means+SEM from three independent experiments. (E, F) Partial colocalization of CpG-STAT3ASO with early endosomes and with RNase H1 after cellular uptake. The confocal microscopy to visualize Cy3-labeled oligonucleotides and (E) early endosomal antigen 1 (EEA1) or (F) RNase H1 in prostate cancer cells (DU145) after 15 min and 4 h of incubation with 250 nM CpG-STAT3ASO Cy3 , respectively. (G) The direct interaction of CpG-STAT3ASO Cy3 with RNAse H1 as measured by in situ proximity ligation assay and confocal microscopy. Cells were incubated with 250 nM CpG-STAT3ASO Cy3 or other labeled control oligonucleotides for 4 h before the analysis; shown are representative images from one of three independent experiments.
    Figure Legend Snippet: CpG-STAT3ASO conjugate design and cell-selective uptake. (A) Single-stranded CpG-STAT3ASO design; subscript “S” = phosphothioated nucleotides; “o” = C3 units of the carbon linker; red = 2’-O-methyl-modified nucleotides. (B, C) The in vitro uptake of CpG-STAT3ASO Alexa488 compared to STAT3ASO Alexa488 by: (B) primary human immune cells (pDC: CD303 + , mDCs: CD1c + , B cells: CD19 + , and T-cells: CD3 + ); (C) mouse dendritic (DC2.4) and macrophage (RAW264.7) cells, and prostate cancer cells (DU145 and RM9). Cells were incubated for 1 h with 500 nM (B-left panel, C) or with various concentrations (B-right panel) of CpG-STAT3ASO Alexa488 and STAT3ASO Alexa488 without any transfection reagents. Oligonucleotide uptake was measured cytofluorimetrically. (D) CpG-STAT3ASO is internalized by prostate cancer cells via scavenger receptor- and clathrin-dependent endocytosis. DU145 cells were pretreated using various endocytosis inhibitors or placed in 4°C for 1 h before incubation with CpG-STAT3ASO Alexa488 (250 nM) or STAT3ASO Alexa488 (750 nM) for another hour. The percentage of Alexa488-positive cells was assessed by flow cytometry; shown are means+SEM from three independent experiments. (E, F) Partial colocalization of CpG-STAT3ASO with early endosomes and with RNase H1 after cellular uptake. The confocal microscopy to visualize Cy3-labeled oligonucleotides and (E) early endosomal antigen 1 (EEA1) or (F) RNase H1 in prostate cancer cells (DU145) after 15 min and 4 h of incubation with 250 nM CpG-STAT3ASO Cy3 , respectively. (G) The direct interaction of CpG-STAT3ASO Cy3 with RNAse H1 as measured by in situ proximity ligation assay and confocal microscopy. Cells were incubated with 250 nM CpG-STAT3ASO Cy3 or other labeled control oligonucleotides for 4 h before the analysis; shown are representative images from one of three independent experiments.

    Techniques Used: Modification, In Vitro, Incubation, Transfection, Flow Cytometry, Cytometry, Confocal Microscopy, Labeling, In Situ, Proximity Ligation Assay

    Systemic administration of CpG-STAT3ASO induces regression of bone-localized mouse prostate tumors in immunocompetent mice. C57BL/6 mice were injected intratibially using RM9 or PPS prostate cancer cells. (A) Biodistribution of systemically injected CpG-STAT3ASO Cy3 and STAT3ASO Cy3 in RM9 tumor-bearing mice. Mice were injected IV using 2.5 mg/kg of either oligonucleotide and euthanized 3 h later. Percentages of Cy3 + T-cells (CD3 + ), macrophages (CD11b + F4/80 + ), DCs (CD11b + CD11c + ) and MDSCs (CD11b + /Gr1 + ) were assessed using flow cytometry in single-cell suspensions of bone marrow or spleen. Results of two independent experiments using a total of 6 mice analyzed individually; means+SEM. (B-C) Systemic administration of CpG-STAT3ASO reduces STAT3 activation in bone-localized prostate tumors and in the tumor-associated immune cells. After tumors were established, mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. After the third treatment, mice were euthanized and pSTAT3 activation was assessed in the tumors using immunohistochemistry (B) and flow cytometry (C) in tumor cells (LSC HI SCC HI CD11b - CD3 - ), MDSCs (CD11b + /Gr1 + ), DCs (CD11b + CD11c + ), and T-cells (CD3 + ). C57BL/6 (D-G) or NSG (H) mice were intratibially injected using RM9-Luc or PPS-Luc prostate cancer cells. After tumors were established mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. (D) Tumor progression was monitored using bioluminescent imaging on the AmiX (Spectral Instruments). (E) Repeated systemic administration of CpG-STAT3ASO induces regression of bone-localized tumors and increases the overall survival of mice. Shown are combined results from two independent experiments ( n = 12 mice/each group). (F) Co-injection of CpG ODN and STAT3ASO fails to reproduce the efficacy of the bi-functional CpG-STAT3ASO conjugate against bone-localized RM9-Luc tumors ( n = 6 mice/each group). (G) Systemic administration of CpG-STAT3ASO induced tumor regression in the bone-localized Pten -deficient tumor model (PPS-Luc). Results were combined from two independent experiments ( n = 12 mice/each group). (H) The antitumor effect of CpG-STAT3ASO depended on the presence of an intact immune system and cannot be achieved in immunodeficient NSG mice ( n = 6 mice/each group).
    Figure Legend Snippet: Systemic administration of CpG-STAT3ASO induces regression of bone-localized mouse prostate tumors in immunocompetent mice. C57BL/6 mice were injected intratibially using RM9 or PPS prostate cancer cells. (A) Biodistribution of systemically injected CpG-STAT3ASO Cy3 and STAT3ASO Cy3 in RM9 tumor-bearing mice. Mice were injected IV using 2.5 mg/kg of either oligonucleotide and euthanized 3 h later. Percentages of Cy3 + T-cells (CD3 + ), macrophages (CD11b + F4/80 + ), DCs (CD11b + CD11c + ) and MDSCs (CD11b + /Gr1 + ) were assessed using flow cytometry in single-cell suspensions of bone marrow or spleen. Results of two independent experiments using a total of 6 mice analyzed individually; means+SEM. (B-C) Systemic administration of CpG-STAT3ASO reduces STAT3 activation in bone-localized prostate tumors and in the tumor-associated immune cells. After tumors were established, mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. After the third treatment, mice were euthanized and pSTAT3 activation was assessed in the tumors using immunohistochemistry (B) and flow cytometry (C) in tumor cells (LSC HI SCC HI CD11b - CD3 - ), MDSCs (CD11b + /Gr1 + ), DCs (CD11b + CD11c + ), and T-cells (CD3 + ). C57BL/6 (D-G) or NSG (H) mice were intratibially injected using RM9-Luc or PPS-Luc prostate cancer cells. After tumors were established mice were treated using IV injections (q2d) of 5 mg/kg of indicated oligonucleotides. (D) Tumor progression was monitored using bioluminescent imaging on the AmiX (Spectral Instruments). (E) Repeated systemic administration of CpG-STAT3ASO induces regression of bone-localized tumors and increases the overall survival of mice. Shown are combined results from two independent experiments ( n = 12 mice/each group). (F) Co-injection of CpG ODN and STAT3ASO fails to reproduce the efficacy of the bi-functional CpG-STAT3ASO conjugate against bone-localized RM9-Luc tumors ( n = 6 mice/each group). (G) Systemic administration of CpG-STAT3ASO induced tumor regression in the bone-localized Pten -deficient tumor model (PPS-Luc). Results were combined from two independent experiments ( n = 12 mice/each group). (H) The antitumor effect of CpG-STAT3ASO depended on the presence of an intact immune system and cannot be achieved in immunodeficient NSG mice ( n = 6 mice/each group).

    Techniques Used: Mouse Assay, Injection, Flow Cytometry, Cytometry, Activation Assay, Immunohistochemistry, Imaging, Functional Assay

    12) Product Images from "Synergistic effect of therapeutic stem cells expressing cytosine deaminase and interferon-beta via apoptotic pathway in the metastatic mouse model of breast cancer"

    Article Title: Synergistic effect of therapeutic stem cells expressing cytosine deaminase and interferon-beta via apoptotic pathway in the metastatic mouse model of breast cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.6719

    Hematoxylin and eosin (H E) staining and PCNA expression level in metastatic breast cancer models Excised brains were fixed in 4% normal formalin and embedded in paraffin. After being cut with a microtome, slides were deparaffined and rehydrated with xylene, ethanol, and tap water. ( A ) H E staining. ( B ) Immunohistochemistry (IHC) staining for PCNA. To confirm the proliferation rate of tumor cells, brain sections were treated with the primary antibody, anti-mouse PCNA. Next, the slide was incubated with biotinylated anti-mouse secondary antibody and stained protein (brown color) was observed following DAB and hematoxylin staining. Dotted line: necrosis or apoptosis area of tumor cells in the brain section. Magnification ×100, ×200.
    Figure Legend Snippet: Hematoxylin and eosin (H E) staining and PCNA expression level in metastatic breast cancer models Excised brains were fixed in 4% normal formalin and embedded in paraffin. After being cut with a microtome, slides were deparaffined and rehydrated with xylene, ethanol, and tap water. ( A ) H E staining. ( B ) Immunohistochemistry (IHC) staining for PCNA. To confirm the proliferation rate of tumor cells, brain sections were treated with the primary antibody, anti-mouse PCNA. Next, the slide was incubated with biotinylated anti-mouse secondary antibody and stained protein (brown color) was observed following DAB and hematoxylin staining. Dotted line: necrosis or apoptosis area of tumor cells in the brain section. Magnification ×100, ×200.

    Techniques Used: Staining, Expressing, Immunohistochemistry, Incubation

    13) Product Images from "New insights into saccular development and vascular formation in lung allografts under the renal capsule"

    Article Title: New insights into saccular development and vascular formation in lung allografts under the renal capsule

    Journal: Mechanisms of development

    doi:

    Lung graft vessels are endogenous to the grafts but are connected to host vessels. (A,B) β-Galactosidase staining of lung graft (A) and kidney (B) sections from wild-type lung rudiments that were grafted into KDR.LacZ mice for 8 days. Host endothelial cells in the glomerular tufts (arrow) and between the tubules (arrowhead) in the host kidney stain positive (blue) for β-galactosidase activity (B), but there are no host endothelial cells in the graft (A). (C) Intravenous perfusion of biotinylated lectin into the host shows that vessels in the 8-day lung grafts are connected to the host circulation, as evidenced by lectin staining of both large vessel (arrow) and capillaries (arrowhead) in the grafts. (D,E) Intravenous perfusion of biotinylated lectin into the host after 4 days (D) and 6 days (E) of grafting shows dot-like areas of lectin staining (arrows) within the mesenchyme of the lung grafts at 4 days, and lectin staining of larger caliber vessels (arrows), but no staining of capillaries in the mesenchyme (arrowheads) at 6 days. Bar: 50 μm.
    Figure Legend Snippet: Lung graft vessels are endogenous to the grafts but are connected to host vessels. (A,B) β-Galactosidase staining of lung graft (A) and kidney (B) sections from wild-type lung rudiments that were grafted into KDR.LacZ mice for 8 days. Host endothelial cells in the glomerular tufts (arrow) and between the tubules (arrowhead) in the host kidney stain positive (blue) for β-galactosidase activity (B), but there are no host endothelial cells in the graft (A). (C) Intravenous perfusion of biotinylated lectin into the host shows that vessels in the 8-day lung grafts are connected to the host circulation, as evidenced by lectin staining of both large vessel (arrow) and capillaries (arrowhead) in the grafts. (D,E) Intravenous perfusion of biotinylated lectin into the host after 4 days (D) and 6 days (E) of grafting shows dot-like areas of lectin staining (arrows) within the mesenchyme of the lung grafts at 4 days, and lectin staining of larger caliber vessels (arrows), but no staining of capillaries in the mesenchyme (arrowheads) at 6 days. Bar: 50 μm.

    Techniques Used: Staining, Mouse Assay, Activity Assay

    14) Product Images from "Inhibition of amyloid-β production by anti-amyloid precursor protein antibodies in primary mouse cortical neurones"

    Article Title: Inhibition of amyloid-β production by anti-amyloid precursor protein antibodies in primary mouse cortical neurones

    Journal: Neuroreport

    doi: 10.1097/WNR.0000000000000055

    Representative examples of immunocytochemistry for NSE, GFAP and 2B3 in mouse primary cortical cultures (A-D). (A) Specific labelling of a large population of wild-type neurones with an anti-NSE antibody (10μg/ml) detected using a biotinylated anti-rabbit antibody (1:500) and Avidin-FITC (1:600); (B) specific labelling of a small population of wild-type glial cells with an anti-GFAP antibody (1μg/ml) detected as above; (C) specific labelling of wild-type cells with 2B3 (10μg/ml) detected using a biotinylated anti-mouse antibody (1:500) and Avidin-FITC (1:600); (D) negative control for 2B3 showing low levels of non-specific staining in wild-type cells incubated in the absence of the primary antibody and detected as above. Similar results were obtained with negative controls for NSE and GFAP (data not shown). Coverslips were mounted with VECTASHIELD mounting medium with DAPI to counter-stain the nuclei. n = 5-7. Scale bar = 25μm.
    Figure Legend Snippet: Representative examples of immunocytochemistry for NSE, GFAP and 2B3 in mouse primary cortical cultures (A-D). (A) Specific labelling of a large population of wild-type neurones with an anti-NSE antibody (10μg/ml) detected using a biotinylated anti-rabbit antibody (1:500) and Avidin-FITC (1:600); (B) specific labelling of a small population of wild-type glial cells with an anti-GFAP antibody (1μg/ml) detected as above; (C) specific labelling of wild-type cells with 2B3 (10μg/ml) detected using a biotinylated anti-mouse antibody (1:500) and Avidin-FITC (1:600); (D) negative control for 2B3 showing low levels of non-specific staining in wild-type cells incubated in the absence of the primary antibody and detected as above. Similar results were obtained with negative controls for NSE and GFAP (data not shown). Coverslips were mounted with VECTASHIELD mounting medium with DAPI to counter-stain the nuclei. n = 5-7. Scale bar = 25μm.

    Techniques Used: Immunocytochemistry, Avidin-Biotin Assay, Negative Control, Staining, Incubation

    15) Product Images from "Cartilage-specific deletion of ephrin-B2 in mice results in early developmental defects and an osteoarthritis-like phenotype during aging in vivo"

    Article Title: Cartilage-specific deletion of ephrin-B2 in mice results in early developmental defects and an osteoarthritis-like phenotype during aging in vivo

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-016-0965-6

    Locomotor phenotype of some ephrin-B2 ( EFNB2 ) Col2 knockout ( KO ) cartilage conditional mice is related to an abnormal corticospinal tract. a - f Brain tracing experiments to visualize the corticospinal path with biotinylated dextran amines ( BDA ) performed in 6-week-old EFNB2 fl/fl (n = 2) and EFNB2 Col2 KO cartilage conditional (n = 3) mice. Representative photomicrographs of the cervical spinal cord of EFNB2 fl/fl ( a , c , d ) and EFNB2 Col2 KO cartilage conditional mice ( b , e , f ) at the level of the fifth cervical vertebra. g Percentage of BDA-labeled fibers in the contralateral and ipsilateral hemispheres. Arrowheads indicate the injection site ( a , b ) and labeled fibers ( c - f ). Scale bars at 100 μm ( a , d ). Data are expressed as the mean ± standard error of the mean and the P values were determined by Mann–Whitney U test
    Figure Legend Snippet: Locomotor phenotype of some ephrin-B2 ( EFNB2 ) Col2 knockout ( KO ) cartilage conditional mice is related to an abnormal corticospinal tract. a - f Brain tracing experiments to visualize the corticospinal path with biotinylated dextran amines ( BDA ) performed in 6-week-old EFNB2 fl/fl (n = 2) and EFNB2 Col2 KO cartilage conditional (n = 3) mice. Representative photomicrographs of the cervical spinal cord of EFNB2 fl/fl ( a , c , d ) and EFNB2 Col2 KO cartilage conditional mice ( b , e , f ) at the level of the fifth cervical vertebra. g Percentage of BDA-labeled fibers in the contralateral and ipsilateral hemispheres. Arrowheads indicate the injection site ( a , b ) and labeled fibers ( c - f ). Scale bars at 100 μm ( a , d ). Data are expressed as the mean ± standard error of the mean and the P values were determined by Mann–Whitney U test

    Techniques Used: Knock-Out, Mouse Assay, Labeling, Injection, MANN-WHITNEY

    16) Product Images from "Epsin is required for Dishevelled stability and Wnt signaling activation in colon cancer development"

    Article Title: Epsin is required for Dishevelled stability and Wnt signaling activation in colon cancer development

    Journal: Nature communications

    doi: 10.1038/ncomms7380

    Epsin interacted with Dvl2 to regulate Wnt receptor signaling independent of receptor-mediated endocytosis (a) Co-immunoprecipitation of endogenous Dvl2, LRP6 and Fzd7 by anti-epsin 1 antibody in Wnt3a-stimulated (100 ng mL −1 , 5 min) HT-29 cells analyzed by Western blotting. (b) Quantification of a , n=8. (c) WT and DKO CECs were stimulated with Wnt3a (100 ng mL −1 ) as indicated then Wnt signaling was analyzed by Western blotting with LRP6 and phospho-LRP6 antibodies. (d) Quantification of phospho-to-total LRP6 fold change in c , n=8. (e) Cell surface of WT and DKO CECs were labeled with cleavable biotin, incubated with Wnt3a at 37°C as indicated then surface biotin was cleaved. Internalized biotinylated-LRP6 was determined by streptavidin bead pull-down and Western blotting. (f) Quantification of e , n=8. All statistical values were calculated using a Student’s t test; P values are indicated. Error bars indicate the mean ± s.e.m.
    Figure Legend Snippet: Epsin interacted with Dvl2 to regulate Wnt receptor signaling independent of receptor-mediated endocytosis (a) Co-immunoprecipitation of endogenous Dvl2, LRP6 and Fzd7 by anti-epsin 1 antibody in Wnt3a-stimulated (100 ng mL −1 , 5 min) HT-29 cells analyzed by Western blotting. (b) Quantification of a , n=8. (c) WT and DKO CECs were stimulated with Wnt3a (100 ng mL −1 ) as indicated then Wnt signaling was analyzed by Western blotting with LRP6 and phospho-LRP6 antibodies. (d) Quantification of phospho-to-total LRP6 fold change in c , n=8. (e) Cell surface of WT and DKO CECs were labeled with cleavable biotin, incubated with Wnt3a at 37°C as indicated then surface biotin was cleaved. Internalized biotinylated-LRP6 was determined by streptavidin bead pull-down and Western blotting. (f) Quantification of e , n=8. All statistical values were calculated using a Student’s t test; P values are indicated. Error bars indicate the mean ± s.e.m.

    Techniques Used: Immunoprecipitation, Western Blot, Labeling, Incubation

    17) Product Images from "Technical Brief: Subretinal injection and electroporation into adult mouse eyes"

    Article Title: Technical Brief: Subretinal injection and electroporation into adult mouse eyes

    Journal: Molecular Vision

    doi:

    Quantum dots in the whole eye after subretinal injection. The mouse was sacrificed 24 h after injection of the quantum dots. This length of time allowed the three blebs to resorb and the neural retina to return to contact with the RPE sheet. Cryosections were collected and counterstained with DAPI (to stain all nuclei blue). A : Shown is a composite of numerous images originally collected with a 20X objective. Images were fused in Photoshop with the Photomerge tool (Adobe Systems Inc., San Jose, CA). The cornea was lanced at clock face position 11, and a small number of quantum dots can be seen in the corneal stroma at the wound site. The retina was punctured between clock face positions 4 and 5. The bright yellow represents intense fluorescence of highly concentrated quantum dots located in the subretinal space. There is a continuous gradient of color from bright yellow to purple, indicating successively less fluorescence of the quantum dots in proportion to their concentration. The quantum dots were confined specifically to the subretinal bleb. The scale bar represents 500 μm. B : A few dots were found in the optic nerve head and interstitial spaces in the optic nerve, as indicated by the red and purple colors. C : A distinctive gradient of color from bright yellow to purple is illustrated from right to left within the confines of the subretinal space. The comparative absence of any color other than blue-stained nuclei in the section suggests that there was no break in the RPE sheet or tear in the neural retina during subretinal injection. D : Close-up reveals quantum dots phagocytosed within the RPE cells. Quantum dots (red) were detected at the level of the RPE cells, and the dots surround the Yo-Pro-1 stained nuclei (green) of the RPE cells. This location indicated that the quantum dots were internalized into the RPE cells. The scale bar represents 50 μm.
    Figure Legend Snippet: Quantum dots in the whole eye after subretinal injection. The mouse was sacrificed 24 h after injection of the quantum dots. This length of time allowed the three blebs to resorb and the neural retina to return to contact with the RPE sheet. Cryosections were collected and counterstained with DAPI (to stain all nuclei blue). A : Shown is a composite of numerous images originally collected with a 20X objective. Images were fused in Photoshop with the Photomerge tool (Adobe Systems Inc., San Jose, CA). The cornea was lanced at clock face position 11, and a small number of quantum dots can be seen in the corneal stroma at the wound site. The retina was punctured between clock face positions 4 and 5. The bright yellow represents intense fluorescence of highly concentrated quantum dots located in the subretinal space. There is a continuous gradient of color from bright yellow to purple, indicating successively less fluorescence of the quantum dots in proportion to their concentration. The quantum dots were confined specifically to the subretinal bleb. The scale bar represents 500 μm. B : A few dots were found in the optic nerve head and interstitial spaces in the optic nerve, as indicated by the red and purple colors. C : A distinctive gradient of color from bright yellow to purple is illustrated from right to left within the confines of the subretinal space. The comparative absence of any color other than blue-stained nuclei in the section suggests that there was no break in the RPE sheet or tear in the neural retina during subretinal injection. D : Close-up reveals quantum dots phagocytosed within the RPE cells. Quantum dots (red) were detected at the level of the RPE cells, and the dots surround the Yo-Pro-1 stained nuclei (green) of the RPE cells. This location indicated that the quantum dots were internalized into the RPE cells. The scale bar represents 50 μm.

    Techniques Used: Injection, Staining, Fluorescence, Concentration Assay

    18) Product Images from "RNase H2 Loss in Murine Astrocytes Results in Cellular Defects Reminiscent of Nucleic Acid-Mediated Autoinflammation"

    Article Title: RNase H2 Loss in Murine Astrocytes Results in Cellular Defects Reminiscent of Nucleic Acid-Mediated Autoinflammation

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00587

    Expression of RNase H2 in the murine central nervous system. (A) Sagittal sections from p21 mouse brains were stained with an RNase H2-specific antiserum followed by a biotinylated secondary antibody, streptavidin-coupled HRP, and developed with the chromogenic substrate 3,3′-Diaminobenzidine (DAB). Note the high expression of RNase H2 in the corpus callosum (cc) and hippocampus, with particularly high expression in the subgranular cell layer (sgcl) of the dentate gyrus (dg). Cornu Ammoni (CA), molecular layer (ml), granular layer (gl). (B) Co-immunofluorescence staining for RNase H2 (red) with the neuronal marker NeuN (green, left panel) and astroglia marker GFAP (green, right panel) shows striking co-localization of RNaseH2 in neuronal cells expressing NeuN in both the cerebral cortex and dg. Scale bar = 150 µm (upper four panels), 10 µm (lower two panels) (C) Co-immunofluorescence staining for RNase H2 shows highest expression of RNase H2 in Ki67-positive proliferating cells. Scale bar = 30 µm.
    Figure Legend Snippet: Expression of RNase H2 in the murine central nervous system. (A) Sagittal sections from p21 mouse brains were stained with an RNase H2-specific antiserum followed by a biotinylated secondary antibody, streptavidin-coupled HRP, and developed with the chromogenic substrate 3,3′-Diaminobenzidine (DAB). Note the high expression of RNase H2 in the corpus callosum (cc) and hippocampus, with particularly high expression in the subgranular cell layer (sgcl) of the dentate gyrus (dg). Cornu Ammoni (CA), molecular layer (ml), granular layer (gl). (B) Co-immunofluorescence staining for RNase H2 (red) with the neuronal marker NeuN (green, left panel) and astroglia marker GFAP (green, right panel) shows striking co-localization of RNaseH2 in neuronal cells expressing NeuN in both the cerebral cortex and dg. Scale bar = 150 µm (upper four panels), 10 µm (lower two panels) (C) Co-immunofluorescence staining for RNase H2 shows highest expression of RNase H2 in Ki67-positive proliferating cells. Scale bar = 30 µm.

    Techniques Used: Expressing, Staining, Immunofluorescence, Marker

    19) Product Images from "Application of protein lysate microarrays to molecular marker verification and quantification"

    Article Title: Application of protein lysate microarrays to molecular marker verification and quantification

    Journal: Proteome Science

    doi: 10.1186/1477-5956-3-9

    Schematic of the different binding steps on LMA. F9 lysates or purified p53 protein, purified primary p53 antibody, biotinylated secondary antibody, and Cy3-labeled streptavidin were printed on spots i, ii, iii and iv on the LMA, respectively. A diagrammatic representation of the different binding steps performed for signal detection on LMA is shown.
    Figure Legend Snippet: Schematic of the different binding steps on LMA. F9 lysates or purified p53 protein, purified primary p53 antibody, biotinylated secondary antibody, and Cy3-labeled streptavidin were printed on spots i, ii, iii and iv on the LMA, respectively. A diagrammatic representation of the different binding steps performed for signal detection on LMA is shown.

    Techniques Used: Binding Assay, Purification, Labeling

    Linearity of different binding steps on LMA. LMA containing serially diluted F9 protein lysates (0.055, 0.11, 0.23, 0.46, 0.92, 1.84, 3.68 and 7.36 ng), purified p53 protein (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), purified primary antibody (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), biotinylated secondary antibody (11.5, 23, 46, 92, 184, 368, 736 and 1472 pg) and Cy3-labeled streptavidin (1.95, 3.9, 7.8, 15.6, 31.2, 62.4, 124.8 and 249.6 ng) were printed. Arrays were probed with p53 antibody followed by binding and labeling steps as described in the Methods. Log transformed (base 2) spotted lysate amount was plotted against log transformed (base 2) signal intensity.
    Figure Legend Snippet: Linearity of different binding steps on LMA. LMA containing serially diluted F9 protein lysates (0.055, 0.11, 0.23, 0.46, 0.92, 1.84, 3.68 and 7.36 ng), purified p53 protein (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), purified primary antibody (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), biotinylated secondary antibody (11.5, 23, 46, 92, 184, 368, 736 and 1472 pg) and Cy3-labeled streptavidin (1.95, 3.9, 7.8, 15.6, 31.2, 62.4, 124.8 and 249.6 ng) were printed. Arrays were probed with p53 antibody followed by binding and labeling steps as described in the Methods. Log transformed (base 2) spotted lysate amount was plotted against log transformed (base 2) signal intensity.

    Techniques Used: Binding Assay, Purification, Labeling, Transformation Assay

    20) Product Images from "Mice Lacking Expression of Secondary Lymphoid Organ Chemokine Have Defects in Lymphocyte Homing and Dendritic Cell Localization "

    Article Title: Mice Lacking Expression of Secondary Lymphoid Organ Chemokine Have Defects in Lymphocyte Homing and Dendritic Cell Localization

    Journal: The Journal of Experimental Medicine

    doi:

    (A) The number of DCs is decreased in LNs of plt mice. LNs of +/+ and plt mice were collected and centrifuged over metrizamide gradients. Cells at the interface were collected, stained with FITC-conjugated anti–mouse I-Ad, normalized to the total number of cells per LN, and analyzed by FACS ® to detect I-A + DCs (top right quadrant). One representative experiment of three is shown. (B) The number of DCs is normal in spleens of plt mice. Spleens of +/+ and plt mice were collected, dissociated with collagenase, and stained with FITC-anti–I-Ad, biotinylated anti-B220, and biotinylated anti-CD3 followed by SA-PerCP, normalized to the total number of cells per spleen, and analyzed by flow cytometry. CD3 − , B220 − gated cells in one representative experiment of three are shown.
    Figure Legend Snippet: (A) The number of DCs is decreased in LNs of plt mice. LNs of +/+ and plt mice were collected and centrifuged over metrizamide gradients. Cells at the interface were collected, stained with FITC-conjugated anti–mouse I-Ad, normalized to the total number of cells per LN, and analyzed by FACS ® to detect I-A + DCs (top right quadrant). One representative experiment of three is shown. (B) The number of DCs is normal in spleens of plt mice. Spleens of +/+ and plt mice were collected, dissociated with collagenase, and stained with FITC-anti–I-Ad, biotinylated anti-B220, and biotinylated anti-CD3 followed by SA-PerCP, normalized to the total number of cells per spleen, and analyzed by flow cytometry. CD3 − , B220 − gated cells in one representative experiment of three are shown.

    Techniques Used: Mouse Assay, Staining, FACS, Flow Cytometry, Cytometry

    Decreased migration of skin DCs to LNs in plt mice after contact sensitization with FITC. (A) The shaved abdomens of +/+ and plt mice were painted with 2 mg FITC. After 24 h draining LNs were removed, dissociated, normalized to the total number of cells per LN, and analyzed by flow cytometry. A decreased number of large FITC + cells (boxed area) can be seen in plt mice. One of eight representative experiments is shown. (B) Representative FACS ® profile showing a marked decrease of CD11c + FITC + cells in plt mice after FITC skin painting. (C) Draining LN cells from FITC-painted mice were partially purified on metrizamide gradients, stained with biotinylated anti–I-Ad followed by SA-PerCP, and analyzed by flow cytometry. Only large FITC + cells (boxed areas in A) are shown. (D) The number of FITC + DCs (boxed areas in A) that accumulate in LNs after skin painting with 2 mg FITC is reduced in plt mice. Numbers represent mean ± SD ( n = 8). (E) Comparison of DC content in contralateral (CLN) and draining (DLN) inguinal LNs in mice painted on one flank with 0.2 mg FITC. Single cell suspensions were prepared from individual LNs, stained with anti–I-Ad and anti-B220, and analyzed by flow cytometry gated on I-A + B220 − cells.
    Figure Legend Snippet: Decreased migration of skin DCs to LNs in plt mice after contact sensitization with FITC. (A) The shaved abdomens of +/+ and plt mice were painted with 2 mg FITC. After 24 h draining LNs were removed, dissociated, normalized to the total number of cells per LN, and analyzed by flow cytometry. A decreased number of large FITC + cells (boxed area) can be seen in plt mice. One of eight representative experiments is shown. (B) Representative FACS ® profile showing a marked decrease of CD11c + FITC + cells in plt mice after FITC skin painting. (C) Draining LN cells from FITC-painted mice were partially purified on metrizamide gradients, stained with biotinylated anti–I-Ad followed by SA-PerCP, and analyzed by flow cytometry. Only large FITC + cells (boxed areas in A) are shown. (D) The number of FITC + DCs (boxed areas in A) that accumulate in LNs after skin painting with 2 mg FITC is reduced in plt mice. Numbers represent mean ± SD ( n = 8). (E) Comparison of DC content in contralateral (CLN) and draining (DLN) inguinal LNs in mice painted on one flank with 0.2 mg FITC. Single cell suspensions were prepared from individual LNs, stained with anti–I-Ad and anti-B220, and analyzed by flow cytometry gated on I-A + B220 − cells.

    Techniques Used: Migration, Mouse Assay, Flow Cytometry, Cytometry, FACS, Purification, Staining

    21) Product Images from "Blood glycemia-modulating effects of melanian snail protein hydrolysates in mice with type II diabetes"

    Article Title: Blood glycemia-modulating effects of melanian snail protein hydrolysates in mice with type II diabetes

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2017.2967

    Histological images of the insulin- and glucagon-immunoreactive cells in the pancreas. Significant increases of insulin and glucagon-immunoreactive cells, and also insulin/glucagon cells were detected in the HFD control mice as compared with the intact control. However, these abnormal increases in insulin and glucagon-immunostained cells and their ratio (insulin/glucagon cells) were significantly normalized by treatment with all test materials, including MPh 250 mg/kg as compared with the HFD control.(A) Intact control: mice supplied normal pellet diet (vehicle control mice); mice administered 10 ml/kg of distilled water orally. (B) HFD control: mice administered 10 ml/kg of distilled water orally with HFD supply. (C) Metformin, mice administered 250 mg/kg of metformin orally with HFD supply. (D) MPh 500, mice administered 500 mg/kg of MPh orally with HFD supply. (E) MPh 250, mice administered 250 mg/kg of MPh orally with HFD supply. (F) MPh 125, mice administered 125 mg/kg of MPh orally with HFD supply. HFD, 45% kcal high-fat diet. MPh, melanian snail ( Semisulcospira libertina ) protein hydrolysates, test material. All images show immunostaining with avidin-biotin-peroxidase complex. Scale bars, 80 µ m.
    Figure Legend Snippet: Histological images of the insulin- and glucagon-immunoreactive cells in the pancreas. Significant increases of insulin and glucagon-immunoreactive cells, and also insulin/glucagon cells were detected in the HFD control mice as compared with the intact control. However, these abnormal increases in insulin and glucagon-immunostained cells and their ratio (insulin/glucagon cells) were significantly normalized by treatment with all test materials, including MPh 250 mg/kg as compared with the HFD control.(A) Intact control: mice supplied normal pellet diet (vehicle control mice); mice administered 10 ml/kg of distilled water orally. (B) HFD control: mice administered 10 ml/kg of distilled water orally with HFD supply. (C) Metformin, mice administered 250 mg/kg of metformin orally with HFD supply. (D) MPh 500, mice administered 500 mg/kg of MPh orally with HFD supply. (E) MPh 250, mice administered 250 mg/kg of MPh orally with HFD supply. (F) MPh 125, mice administered 125 mg/kg of MPh orally with HFD supply. HFD, 45% kcal high-fat diet. MPh, melanian snail ( Semisulcospira libertina ) protein hydrolysates, test material. All images show immunostaining with avidin-biotin-peroxidase complex. Scale bars, 80 µ m.

    Techniques Used: Mouse Assay, Immunostaining, Avidin-Biotin Assay

    22) Product Images from "Improved antifouling properties and selective biofunctionalization of stainless steel by employing heterobifunctional silane-polyethylene glycol overlayers and avidin-biotin technology"

    Article Title: Improved antifouling properties and selective biofunctionalization of stainless steel by employing heterobifunctional silane-polyethylene glycol overlayers and avidin-biotin technology

    Journal: Scientific Reports

    doi: 10.1038/srep29324

    Biofunctionalization of SS-SPB. ( a ) Schematic illustration of the immobilization of bAP on SPB-modified SS via avidin-biotin bridge. ( b ) Spectrophotometric detection of the activity of surface-immobilized bAP. In the negative control, avidin binding sites had been blocked with free biotin to prevent selective attachment. ( c,d ) AFM images showing avidin functionalized SS-SPB. Scale bars 200 nm and 1 μm, respectively. ( e,f ) SS-SPB surface without avidin functionalization. Scale bars as in ( c,d ).
    Figure Legend Snippet: Biofunctionalization of SS-SPB. ( a ) Schematic illustration of the immobilization of bAP on SPB-modified SS via avidin-biotin bridge. ( b ) Spectrophotometric detection of the activity of surface-immobilized bAP. In the negative control, avidin binding sites had been blocked with free biotin to prevent selective attachment. ( c,d ) AFM images showing avidin functionalized SS-SPB. Scale bars 200 nm and 1 μm, respectively. ( e,f ) SS-SPB surface without avidin functionalization. Scale bars as in ( c,d ).

    Techniques Used: Modification, Avidin-Biotin Assay, Activity Assay, Negative Control, Binding Assay

    Schematic illustration of silane-PEG-modified SS. ( a ) SPC overlayer resists non-specific biofouling of both bacteria and proteins. ( b ) SPB overlayer enables selective functionalization of SS via avidin-biotin bridges with bAP without compromising the antifouling properties.
    Figure Legend Snippet: Schematic illustration of silane-PEG-modified SS. ( a ) SPC overlayer resists non-specific biofouling of both bacteria and proteins. ( b ) SPB overlayer enables selective functionalization of SS via avidin-biotin bridges with bAP without compromising the antifouling properties.

    Techniques Used: Modification, Avidin-Biotin Assay

    23) Product Images from "Preserved Calretinin Interneurons in an App Model of Alzheimer’s Disease Disrupt Hippocampal Inhibition via Upregulated P2Y1 Purinoreceptors"

    Article Title: Preserved Calretinin Interneurons in an App Model of Alzheimer’s Disease Disrupt Hippocampal Inhibition via Upregulated P2Y1 Purinoreceptors

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhz165

    Age-dependent phenotypical changes in the App NL-F/NL-F model of AD. ( A , C , E ) Z -stack images from confocal microscopy illustrating the expression of GFAP (for reactive astrocytes), CD68 (for microglia), and Aβ (all in red, secondary antibody Texas Red) together with DAPI staining for nuclei (in blue) in 12-month age-matched wild-type and App NL-F/NL-F mice, respectively. Similarly, bright-field images of tissue immunostained with biotinylated antibodies show conglomerates of GFAP, CD68, and Aβ in the same animals. Both immunofluorescence and immunoperoxidase-stained images taken at ×20 magnification (larger images, scale bar = 50 μm) and ×63 magnification (inserts, scale bar = 20 μm). ( B , D , F ) Analysis of GFAP, CD68, and Aβ from immunoperoxidase-stained tissue. Significant differences in the three markers of AD were seen between wild-type and App NL-F/NL-F mice only at 9–18 months and when comparing quantification at 9–18 months with the other two age cohorts. ( G , I ) Age-dependent accumulation of Aβ in selective subtypes of interneurons in hippocampal CA1. Aβ colocalization was found at significantly higher levels in SST and CCK cells (indicated by arrows), but not in calretinin (CR) cells in the same animals at 12 months (scale = 20 μm). ( J ) Quantification of colocalization of Aβ with either CCK, SST, or calretinin cells. A two-way ANOVA was performed with pairwise comparisons corrected for multiple comparisons (α = 0.05), with either post hoc Sidak’s test or Tukey’s test for multiple comparisons. * P
    Figure Legend Snippet: Age-dependent phenotypical changes in the App NL-F/NL-F model of AD. ( A , C , E ) Z -stack images from confocal microscopy illustrating the expression of GFAP (for reactive astrocytes), CD68 (for microglia), and Aβ (all in red, secondary antibody Texas Red) together with DAPI staining for nuclei (in blue) in 12-month age-matched wild-type and App NL-F/NL-F mice, respectively. Similarly, bright-field images of tissue immunostained with biotinylated antibodies show conglomerates of GFAP, CD68, and Aβ in the same animals. Both immunofluorescence and immunoperoxidase-stained images taken at ×20 magnification (larger images, scale bar = 50 μm) and ×63 magnification (inserts, scale bar = 20 μm). ( B , D , F ) Analysis of GFAP, CD68, and Aβ from immunoperoxidase-stained tissue. Significant differences in the three markers of AD were seen between wild-type and App NL-F/NL-F mice only at 9–18 months and when comparing quantification at 9–18 months with the other two age cohorts. ( G , I ) Age-dependent accumulation of Aβ in selective subtypes of interneurons in hippocampal CA1. Aβ colocalization was found at significantly higher levels in SST and CCK cells (indicated by arrows), but not in calretinin (CR) cells in the same animals at 12 months (scale = 20 μm). ( J ) Quantification of colocalization of Aβ with either CCK, SST, or calretinin cells. A two-way ANOVA was performed with pairwise comparisons corrected for multiple comparisons (α = 0.05), with either post hoc Sidak’s test or Tukey’s test for multiple comparisons. * P

    Techniques Used: Confocal Microscopy, Expressing, Staining, Mouse Assay, Immunofluorescence

    24) Product Images from "Clinical significance of BRCA1 and BRCA2 mRNA and protein expression in patients with sporadic gastric cancer"

    Article Title: Clinical significance of BRCA1 and BRCA2 mRNA and protein expression in patients with sporadic gastric cancer

    Journal: Oncology Letters

    doi: 10.3892/ol.2019.10132

    Immunohistochemistry of BRCA1 and BRCA2 protein expression in gastric cancer tissues. (A) Strong positive BRCA1 staining in the cytoplasm and nuclei, and (B) positive BRCA2 staining in the cytoplasm of gastric adenocarcinoma cells. (C) Negative BRCA1 staining in the cytoplasm and nuclei, and (D) negative BRCA2 staining in the nucleus and cytoplasm of gastric adenocarcinoma cells. (E) Positive BRCA1 staining in the cytoplasm, and (F) positive BRCA2 staining in the cytoplasm of chronic gastritis cells. Magnification, ×400 (scale bar, 50 µm). BRCA1/2, breast cancer type 1 and 2 susceptibility protein.
    Figure Legend Snippet: Immunohistochemistry of BRCA1 and BRCA2 protein expression in gastric cancer tissues. (A) Strong positive BRCA1 staining in the cytoplasm and nuclei, and (B) positive BRCA2 staining in the cytoplasm of gastric adenocarcinoma cells. (C) Negative BRCA1 staining in the cytoplasm and nuclei, and (D) negative BRCA2 staining in the nucleus and cytoplasm of gastric adenocarcinoma cells. (E) Positive BRCA1 staining in the cytoplasm, and (F) positive BRCA2 staining in the cytoplasm of chronic gastritis cells. Magnification, ×400 (scale bar, 50 µm). BRCA1/2, breast cancer type 1 and 2 susceptibility protein.

    Techniques Used: Immunohistochemistry, Expressing, Staining

    (A) Diagram illustrating the distribution of patients according to the mean value (3.90) of the BRCA1 mRNA expression level into low (0.06–3.84; mean, 1.27) and high (4.10–33.26; mean, 9.53) groups (outliners: Open circle, > 1.5× interquartile range; asterisk, > 3× interquartile range). (B) Diagram illustrating the distribution of patients according to the mean value (1.00) of the BRCA1 mRNA expression level into low (0.04–1.00; mean, 0.38) and high (1.10–25.42; mean, 4.19) groups (outliners: Open circle, > 1.5X interquartile range). BRCA1/2 , BRCA1/2 DNA repair associated.
    Figure Legend Snippet: (A) Diagram illustrating the distribution of patients according to the mean value (3.90) of the BRCA1 mRNA expression level into low (0.06–3.84; mean, 1.27) and high (4.10–33.26; mean, 9.53) groups (outliners: Open circle, > 1.5× interquartile range; asterisk, > 3× interquartile range). (B) Diagram illustrating the distribution of patients according to the mean value (1.00) of the BRCA1 mRNA expression level into low (0.04–1.00; mean, 0.38) and high (1.10–25.42; mean, 4.19) groups (outliners: Open circle, > 1.5X interquartile range). BRCA1/2 , BRCA1/2 DNA repair associated.

    Techniques Used: Expressing

    Kaplan-Meier survival analysis of patients with gastric cancer. Overall survival according to (A) BRCA1 IHC and (B) BRCA2 IHC. Disease free survival according to (C) BRCA1 IHC and, (D) BRCA2 IHC. Blue line; negative expression of BRCA1/BRCA2 protein by IHC, red line; positive expression of BRCA1/BRCA2 protein by IHC. BRCA1/2, breast cancer type 1 and 2 susceptibility protein; IHC, immunohistochemistry.
    Figure Legend Snippet: Kaplan-Meier survival analysis of patients with gastric cancer. Overall survival according to (A) BRCA1 IHC and (B) BRCA2 IHC. Disease free survival according to (C) BRCA1 IHC and, (D) BRCA2 IHC. Blue line; negative expression of BRCA1/BRCA2 protein by IHC, red line; positive expression of BRCA1/BRCA2 protein by IHC. BRCA1/2, breast cancer type 1 and 2 susceptibility protein; IHC, immunohistochemistry.

    Techniques Used: Immunohistochemistry, Expressing

    25) Product Images from "Intratumoral Wnt2B expression affects tumor proliferation and survival in malignant pleural mesothelioma patients"

    Article Title: Intratumoral Wnt2B expression affects tumor proliferation and survival in malignant pleural mesothelioma patients

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2012.511

    Immunohistochemical staining of human MPM tissue using the avidin-biotin-peroxidase complex procedure. (A) A Wnt1-high tumor. A tumor with positive expression of (B) Wnt2B, (C) nuclear survivin and (D) c-Myc. A tumor with negative expression of (E) Wnt2B, (F) nuclear survivin and (G) c-Myc. (H) A Wnt5A-high tumor. Original magnification, ×200.
    Figure Legend Snippet: Immunohistochemical staining of human MPM tissue using the avidin-biotin-peroxidase complex procedure. (A) A Wnt1-high tumor. A tumor with positive expression of (B) Wnt2B, (C) nuclear survivin and (D) c-Myc. A tumor with negative expression of (E) Wnt2B, (F) nuclear survivin and (G) c-Myc. (H) A Wnt5A-high tumor. Original magnification, ×200.

    Techniques Used: Immunohistochemistry, Staining, Avidin-Biotin Assay, Expressing

    26) Product Images from "Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice"

    Article Title: Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4720-05.2006

    D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a biotinylated donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.
    Figure Legend Snippet: D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a biotinylated donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.

    Techniques Used: Immunocytochemistry, Mouse Assay, Knock-Out

    27) Product Images from "Antigen Display, T-Cell Activation, and Immune Evasion during Acute and Chronic Ehrlichiosis "

    Article Title: Antigen Display, T-Cell Activation, and Immune Evasion during Acute and Chronic Ehrlichiosis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01433-08

    DCs display antigen and harbor E. muris . DCs were purified from pooled day 5-infected spleens and LNs ( n = 3), enriched by magnetic bead positive selection, and then purified by flow-cytometric cell sorting (95 to 99% purity was obtained). (a) Representative dot plots demonstrating the homogeneity of each of the purified DC populations. (b) OMP-19 107-122 antigen presentation was measured in cultures containing the DCs, purified as described for panel a, from mock-infected mice (mock) or from spleen and LNs from mice on day 5 postinfection. The data are representative of two independent experiments of similar design; the average frequencies of IFN-γ-producing T cells were 43.7 and 13.6% for spleen and LNs, respectively. (c) DCs from mice on day 5 postinfection were purified by flow-cytometric cell sorting as shown in panel a and were stained with antibodies that recognize E. muris (Ec18.1) and CD11c. The staining for E. muris was performed using biotinylated Ec18.1. The cells first were stained using streptavidin-conjugated Alexafluor-594 (pseudocolored cyan in the figure), permeabilized with 0.2% saponin, blocked, and stained again with streptavidin-conjugated Alexafluor-488 (shown in green). Thus, the cyan-colored bacteria are surface-associated bacteria, and the green bacteria represent all of the bacteria associated with the host cell. The merged image, which also includes CD11c staining (in red), is shown in the panels at the right. Nuclei were counterstained with 4′,6′-diamidino-2-phenylindole (blue in nuclei). The top and bottom rows show two representative fields of cells.
    Figure Legend Snippet: DCs display antigen and harbor E. muris . DCs were purified from pooled day 5-infected spleens and LNs ( n = 3), enriched by magnetic bead positive selection, and then purified by flow-cytometric cell sorting (95 to 99% purity was obtained). (a) Representative dot plots demonstrating the homogeneity of each of the purified DC populations. (b) OMP-19 107-122 antigen presentation was measured in cultures containing the DCs, purified as described for panel a, from mock-infected mice (mock) or from spleen and LNs from mice on day 5 postinfection. The data are representative of two independent experiments of similar design; the average frequencies of IFN-γ-producing T cells were 43.7 and 13.6% for spleen and LNs, respectively. (c) DCs from mice on day 5 postinfection were purified by flow-cytometric cell sorting as shown in panel a and were stained with antibodies that recognize E. muris (Ec18.1) and CD11c. The staining for E. muris was performed using biotinylated Ec18.1. The cells first were stained using streptavidin-conjugated Alexafluor-594 (pseudocolored cyan in the figure), permeabilized with 0.2% saponin, blocked, and stained again with streptavidin-conjugated Alexafluor-488 (shown in green). Thus, the cyan-colored bacteria are surface-associated bacteria, and the green bacteria represent all of the bacteria associated with the host cell. The merged image, which also includes CD11c staining (in red), is shown in the panels at the right. Nuclei were counterstained with 4′,6′-diamidino-2-phenylindole (blue in nuclei). The top and bottom rows show two representative fields of cells.

    Techniques Used: Purification, Infection, Selection, Flow Cytometry, FACS, Mouse Assay, Staining

    28) Product Images from "Epsin is required for Dishevelled stability and Wnt signaling activation in colon cancer development"

    Article Title: Epsin is required for Dishevelled stability and Wnt signaling activation in colon cancer development

    Journal: Nature communications

    doi: 10.1038/ncomms7380

    Epsin interacted with Dvl2 to regulate Wnt receptor signaling independent of receptor-mediated endocytosis (a) Co-immunoprecipitation of endogenous Dvl2, LRP6 and Fzd7 by anti-epsin 1 antibody in Wnt3a-stimulated (100 ng mL −1 , 5 min) HT-29 cells analyzed by Western blotting. (b) Quantification of a , n=8. (c) WT and DKO CECs were stimulated with Wnt3a (100 ng mL −1 ) as indicated then Wnt signaling was analyzed by Western blotting with LRP6 and phospho-LRP6 antibodies. (d) Quantification of phospho-to-total LRP6 fold change in c , n=8. (e) Cell surface of WT and DKO CECs were labeled with cleavable biotin, incubated with Wnt3a at 37°C as indicated then surface biotin was cleaved. Internalized biotinylated-LRP6 was determined by streptavidin bead pull-down and Western blotting. (f) Quantification of e , n=8. All statistical values were calculated using a Student’s t test; P values are indicated. Error bars indicate the mean ± s.e.m.
    Figure Legend Snippet: Epsin interacted with Dvl2 to regulate Wnt receptor signaling independent of receptor-mediated endocytosis (a) Co-immunoprecipitation of endogenous Dvl2, LRP6 and Fzd7 by anti-epsin 1 antibody in Wnt3a-stimulated (100 ng mL −1 , 5 min) HT-29 cells analyzed by Western blotting. (b) Quantification of a , n=8. (c) WT and DKO CECs were stimulated with Wnt3a (100 ng mL −1 ) as indicated then Wnt signaling was analyzed by Western blotting with LRP6 and phospho-LRP6 antibodies. (d) Quantification of phospho-to-total LRP6 fold change in c , n=8. (e) Cell surface of WT and DKO CECs were labeled with cleavable biotin, incubated with Wnt3a at 37°C as indicated then surface biotin was cleaved. Internalized biotinylated-LRP6 was determined by streptavidin bead pull-down and Western blotting. (f) Quantification of e , n=8. All statistical values were calculated using a Student’s t test; P values are indicated. Error bars indicate the mean ± s.e.m.

    Techniques Used: Immunoprecipitation, Western Blot, Labeling, Incubation

    29) Product Images from "Beneficial effects of dried pomegranate juice concentrated powder on ultraviolet B-induced skin photoaging in hairless mice"

    Article Title: Beneficial effects of dried pomegranate juice concentrated powder on ultraviolet B-induced skin photoaging in hairless mice

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2017.4626

    Representative immunohistochemistrical images of dorsal back skin tissues, taken from unexposed intact or UVB-exposed hairless mice. (A) Intact vehicle control; (B) UVB control; (C) UVB-exposed mice treated with 2 ml/kg PCS mice; (D) UVB-exposed mice treated with 100 mg/kg PCP; (E) UVB-exposed mice treated with 200 mg/kg PCP; (F) UVB-exposed mice treated with 400 mg/kg PCP. All avidin-biotin complex immunostaining. Scale bars=50 µm. UVB, ultraviolet B; 4-HNE, 4-hydroxynonenal; PARP, cleaved poly(ADP-ribose) polymerase; MMP-9, matrix metalloproteinase 9; PCP, dried pomegranate juice concentrated powder; PCS, pomegranate juice concentrated solution.
    Figure Legend Snippet: Representative immunohistochemistrical images of dorsal back skin tissues, taken from unexposed intact or UVB-exposed hairless mice. (A) Intact vehicle control; (B) UVB control; (C) UVB-exposed mice treated with 2 ml/kg PCS mice; (D) UVB-exposed mice treated with 100 mg/kg PCP; (E) UVB-exposed mice treated with 200 mg/kg PCP; (F) UVB-exposed mice treated with 400 mg/kg PCP. All avidin-biotin complex immunostaining. Scale bars=50 µm. UVB, ultraviolet B; 4-HNE, 4-hydroxynonenal; PARP, cleaved poly(ADP-ribose) polymerase; MMP-9, matrix metalloproteinase 9; PCP, dried pomegranate juice concentrated powder; PCS, pomegranate juice concentrated solution.

    Techniques Used: Mouse Assay, Avidin-Biotin Assay, Immunostaining

    30) Product Images from "A prokaryotic viral sequence is expressed and conserved in mammalian brain"

    Article Title: A prokaryotic viral sequence is expressed and conserved in mammalian brain

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1706110114

    Spx1 protein localization in mouse GT1 neuron derived cells ( A–E ) and spinal cord ( F ). ( A–E ) Weak but positive (red) staining in cells without GdnHCl unmasking ( A ), and more intense staining in 2 and 3 M GdnHCl-treated cells ( B and C ). Staining was completely blocked by the spx1 peptide ( D ), as shown in parallel stained cells first exposed to 3 M GdnHCl. (Scale bar in A , 100 μm, for A–D.) Higher magnification after 2 M GdnHCl treatment shows the most intense spx1 signal in perinuclear region and some fibril-like elements in the uppermost GT1 cell ( E ). Staining patterns were the same as representatively shown here in uninfected ( A – D ) and FU-CJD-infected cells ( E ). ( F ) Murine insulin-producing pancreatic islet cells (at arrow) are intensely stained, whereas exocrine pancreatic cells have no signal. ( G – I ) Slices containing five representative coronal brain sections with a spleen section at bottom (spl) treated in parallel. Omitting spx primary anitbody gave no staining in brain or spleen ( G ), showing that biotinylated secondary antibody, strepavidin alkaline phosphatase, and detector development reagents give no substantial signal. In contrast, strong positive staining is seen when the primary spx1 antibody is included ( H ). Peptide blocking again, as in GT1 cells, abolished staining of both tissue types ( I ). Mouse spinal cord ( J ) in 5-μm sections shows intense spx1 signal in synaptic boutons (arrows), many of which innervate anterior motor horn neuron dendrites and cell bodies. Nuclei lack signal (also confirmed in noncounterstained section), no labeled boutons are seen on vessels (v), and a myelinated axon (m) is also not labeled. (Scale bar, 20 μm.)
    Figure Legend Snippet: Spx1 protein localization in mouse GT1 neuron derived cells ( A–E ) and spinal cord ( F ). ( A–E ) Weak but positive (red) staining in cells without GdnHCl unmasking ( A ), and more intense staining in 2 and 3 M GdnHCl-treated cells ( B and C ). Staining was completely blocked by the spx1 peptide ( D ), as shown in parallel stained cells first exposed to 3 M GdnHCl. (Scale bar in A , 100 μm, for A–D.) Higher magnification after 2 M GdnHCl treatment shows the most intense spx1 signal in perinuclear region and some fibril-like elements in the uppermost GT1 cell ( E ). Staining patterns were the same as representatively shown here in uninfected ( A – D ) and FU-CJD-infected cells ( E ). ( F ) Murine insulin-producing pancreatic islet cells (at arrow) are intensely stained, whereas exocrine pancreatic cells have no signal. ( G – I ) Slices containing five representative coronal brain sections with a spleen section at bottom (spl) treated in parallel. Omitting spx primary anitbody gave no staining in brain or spleen ( G ), showing that biotinylated secondary antibody, strepavidin alkaline phosphatase, and detector development reagents give no substantial signal. In contrast, strong positive staining is seen when the primary spx1 antibody is included ( H ). Peptide blocking again, as in GT1 cells, abolished staining of both tissue types ( I ). Mouse spinal cord ( J ) in 5-μm sections shows intense spx1 signal in synaptic boutons (arrows), many of which innervate anterior motor horn neuron dendrites and cell bodies. Nuclei lack signal (also confirmed in noncounterstained section), no labeled boutons are seen on vessels (v), and a myelinated axon (m) is also not labeled. (Scale bar, 20 μm.)

    Techniques Used: Derivative Assay, Staining, Infection, Blocking Assay, Labeling

    31) Product Images from "Characterization of Early Pathological Tau Conformations and Phosphorylation in Chronic Traumatic Encephalopathy"

    Article Title: Characterization of Early Pathological Tau Conformations and Phosphorylation in Chronic Traumatic Encephalopathy

    Journal: Journal of neuropathology and experimental neurology

    doi: 10.1093/jnen/nlv001

    Early, pretangle tau pathology markers in cortical neurons in chronic traumatic encephalopathy (CTE). (A–F) Within the superior frontal gyrus, antibodies TNT1 (A, B), TOC1 (C, D) , and pS422 (E, F) all label early, pretangle pathology in pyramidal
    Figure Legend Snippet: Early, pretangle tau pathology markers in cortical neurons in chronic traumatic encephalopathy (CTE). (A–F) Within the superior frontal gyrus, antibodies TNT1 (A, B), TOC1 (C, D) , and pS422 (E, F) all label early, pretangle pathology in pyramidal

    Techniques Used:

    Colocalization of TNT1 tau, TOC1 tau, and pS422 tau in astrocytes in chronic traumatic encephalopathy (CTE). (A–D) Astrocytic tau pathology in the superior frontal gyrus area stained with TNT1 (A) , TOC1 (B) , and pS422 (C) antibodies; merged image
    Figure Legend Snippet: Colocalization of TNT1 tau, TOC1 tau, and pS422 tau in astrocytes in chronic traumatic encephalopathy (CTE). (A–D) Astrocytic tau pathology in the superior frontal gyrus area stained with TNT1 (A) , TOC1 (B) , and pS422 (C) antibodies; merged image

    Techniques Used: Staining

    Colocalization of TNT1, TOC1, and pS422 tau in neuronal pathology of the chronic traumatic encephalopathy (CTE) brain. (A–D) Confocal microscopy was used to image multilabel immunofluorescence staining for TNT1 (A) , TOC1 (B) , and pS422 (C) tau
    Figure Legend Snippet: Colocalization of TNT1, TOC1, and pS422 tau in neuronal pathology of the chronic traumatic encephalopathy (CTE) brain. (A–D) Confocal microscopy was used to image multilabel immunofluorescence staining for TNT1 (A) , TOC1 (B) , and pS422 (C) tau

    Techniques Used: Confocal Microscopy, Immunofluorescence, Staining

    Axonal pathology in the white matter in chronic traumatic encephalopathy (CTE). (A–C) In CTE, axonal thread-like neurites were immunolabeled with TNT1 (A) , TOC1 (B) , and pS422 (C) antibodies. These data suggest that phosphatase-activating domain
    Figure Legend Snippet: Axonal pathology in the white matter in chronic traumatic encephalopathy (CTE). (A–C) In CTE, axonal thread-like neurites were immunolabeled with TNT1 (A) , TOC1 (B) , and pS422 (C) antibodies. These data suggest that phosphatase-activating domain

    Techniques Used: Immunolabeling

    Antibodies to TNT1, TOC1, and pS422 label tau pathology in chronic traumatic encephalopathy (CTE). ( A–D ) Perivascular (v-vessel, dashed outline) tau pathology is immunoreactive for TNT1 ( A ), TOC1 ( B ), and pS422 ( C ). Note the sparse presence of
    Figure Legend Snippet: Antibodies to TNT1, TOC1, and pS422 label tau pathology in chronic traumatic encephalopathy (CTE). ( A–D ) Perivascular (v-vessel, dashed outline) tau pathology is immunoreactive for TNT1 ( A ), TOC1 ( B ), and pS422 ( C ). Note the sparse presence of

    Techniques Used:

    Astrocytic tau pathology in the superior frontal gyrus in chronic traumatic encephalopathy (CTE). (A–C) Clusters of tau-immunoreactive astrocytes in the gray matter immunolabeled with all 3 early tau markers: TNT1 (A) , TOC1 (B) , and pS422 (C)
    Figure Legend Snippet: Astrocytic tau pathology in the superior frontal gyrus in chronic traumatic encephalopathy (CTE). (A–C) Clusters of tau-immunoreactive astrocytes in the gray matter immunolabeled with all 3 early tau markers: TNT1 (A) , TOC1 (B) , and pS422 (C)

    Techniques Used: Immunolabeling

    32) Product Images from "Interfacial Polymerization for Colorimetric Labeling of Protein Expression in Cells"

    Article Title: Interfacial Polymerization for Colorimetric Labeling of Protein Expression in Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0115630

    Compatibility of Polymer Dye Labeling with Vectashield mounting medium. Polymer Dye Labeling of nuclear pore complex imaged (a) dry or (b) in Vectashield hardset mounting medium. Polymer coated nuclei without Evans Blue dye imaged (c) dry or (d) in Vectashield hardset mounting medium. Scale bars are 50 µm.
    Figure Legend Snippet: Compatibility of Polymer Dye Labeling with Vectashield mounting medium. Polymer Dye Labeling of nuclear pore complex imaged (a) dry or (b) in Vectashield hardset mounting medium. Polymer coated nuclei without Evans Blue dye imaged (c) dry or (d) in Vectashield hardset mounting medium. Scale bars are 50 µm.

    Techniques Used: Labeling

    33) Product Images from "Postnatal Generation of Neurons in the Ventrobasal Nucleus of the Rat Thalamus"

    Article Title: Postnatal Generation of Neurons in the Ventrobasal Nucleus of the Rat Thalamus

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1194-07.2007

    Phenotype of cycling cells. Double immunofluorescent labeling studies were used to determine that some of the cycling cells (labeled by an injection of BrdU on P6) expressed neuron-specific markers ≥6 d after injection. A–C , Confocal images were obtained with optical section thickness of 1.0 μm. Sections processed for NeuN ( A ; green arrows) and BrdU ( B ; red arrows) immunohistochemistry are shown. Many BrdU-positive cells were NeuN positive ( C ; yellow arrows). D–F , Images of double labeling with anti-NeuN ( D ) and anti-BrdU ( E ) antibodies. F shows colocalization of the labels in a merged image flanked by images at orthogonal planes through the cell. The orthogonal images were compiled from stacks of 1.0 μm optical images. G–K , Images of labeling with anti-HuC/D antibody ( G ) and anti-BrdU antibody ( H ). The merged images show colocalization of the labels. I , J , A z-series of images was taken through the section shown in G . Individual images were taken at planes that highlighted a nuclear profile of cell 1 ( I ) and cell 2 ( J as supplemental material). L , Animals given injections of BrdU on P6 were given an intracerebral injection of biotinylated dextran on P24 (box inset, I′ as supplemental material). Scale bars: A–C , L , 50 μm; D–K , 10 μm.
    Figure Legend Snippet: Phenotype of cycling cells. Double immunofluorescent labeling studies were used to determine that some of the cycling cells (labeled by an injection of BrdU on P6) expressed neuron-specific markers ≥6 d after injection. A–C , Confocal images were obtained with optical section thickness of 1.0 μm. Sections processed for NeuN ( A ; green arrows) and BrdU ( B ; red arrows) immunohistochemistry are shown. Many BrdU-positive cells were NeuN positive ( C ; yellow arrows). D–F , Images of double labeling with anti-NeuN ( D ) and anti-BrdU ( E ) antibodies. F shows colocalization of the labels in a merged image flanked by images at orthogonal planes through the cell. The orthogonal images were compiled from stacks of 1.0 μm optical images. G–K , Images of labeling with anti-HuC/D antibody ( G ) and anti-BrdU antibody ( H ). The merged images show colocalization of the labels. I , J , A z-series of images was taken through the section shown in G . Individual images were taken at planes that highlighted a nuclear profile of cell 1 ( I ) and cell 2 ( J as supplemental material). L , Animals given injections of BrdU on P6 were given an intracerebral injection of biotinylated dextran on P24 (box inset, I′ as supplemental material). Scale bars: A–C , L , 50 μm; D–K , 10 μm.

    Techniques Used: Labeling, Injection, Immunohistochemistry

    34) Product Images from "Oxidized Phospholipids are Proinflammatory and Proatherogenic in Hypercholesterolemic Mice"

    Article Title: Oxidized Phospholipids are Proinflammatory and Proatherogenic in Hypercholesterolemic Mice

    Journal: Nature

    doi: 10.1038/s41586-018-0198-8

    E06-scFv decreases early aortic valve stenosis, hepatic steatosis, and systemic inflammation a, b, Ldlr −/− (n=11) and Ldlr −/− / E06-scFv (n=10) mice were fed HCD for 15 months and prospectively examined at 3 time points for aortic valve hemodynamics. a , Mean pressure gradients across the aortic valve, determined by Doppler echocardiography. At 12 months there was a 49% lower mean gradient in the Ldlr −/− / E06-scFv mice (2.4±1.9mmHg vs. 4.8±2.4mmHg, mean±SD, p = 0.026, Ldlr −/− / E06-scFv (n=10) and Ldlr −/− (n=9). c, d , Calcification in aortic valve leaflets was determined by von Kossa staining of serial aortic valve sections and AUC compared. AV calcium was reduced in Ldlr −/− / E06-scFv mice by 41.5% (p=0.045, one-tailed-t-test, Ldlr −/− / E06-scFv (n=9) and Ldlr −/− (n=8)). e , Survival of mice used in AV hemodynamic study over 15 months. f , Hepatic cholesterol and triglyceride (TG) levels were reduced by 42% and 47% respectively in Ldlr −/− /E06-scFv mice, Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice g , Livers of mice fed HCD for 16-wks were immunostained with biotinylated E06 IgM (brown) and compared to chow-fed C57BL/6 mice. Shown are representative photomicrographs, representative of 7 Ldlr −/− , 7 Ldlr −/− / E06-scFv and 3 WT (C57BL/6) mice. h , Plasma serum amyloid A (SAA) was decreased 32% in HC fed Ldlr −/− / E06-scFv mice ( Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice).
    Figure Legend Snippet: E06-scFv decreases early aortic valve stenosis, hepatic steatosis, and systemic inflammation a, b, Ldlr −/− (n=11) and Ldlr −/− / E06-scFv (n=10) mice were fed HCD for 15 months and prospectively examined at 3 time points for aortic valve hemodynamics. a , Mean pressure gradients across the aortic valve, determined by Doppler echocardiography. At 12 months there was a 49% lower mean gradient in the Ldlr −/− / E06-scFv mice (2.4±1.9mmHg vs. 4.8±2.4mmHg, mean±SD, p = 0.026, Ldlr −/− / E06-scFv (n=10) and Ldlr −/− (n=9). c, d , Calcification in aortic valve leaflets was determined by von Kossa staining of serial aortic valve sections and AUC compared. AV calcium was reduced in Ldlr −/− / E06-scFv mice by 41.5% (p=0.045, one-tailed-t-test, Ldlr −/− / E06-scFv (n=9) and Ldlr −/− (n=8)). e , Survival of mice used in AV hemodynamic study over 15 months. f , Hepatic cholesterol and triglyceride (TG) levels were reduced by 42% and 47% respectively in Ldlr −/− /E06-scFv mice, Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice g , Livers of mice fed HCD for 16-wks were immunostained with biotinylated E06 IgM (brown) and compared to chow-fed C57BL/6 mice. Shown are representative photomicrographs, representative of 7 Ldlr −/− , 7 Ldlr −/− / E06-scFv and 3 WT (C57BL/6) mice. h , Plasma serum amyloid A (SAA) was decreased 32% in HC fed Ldlr −/− / E06-scFv mice ( Ldlr −/− (n=10) and Ldlr −/− / E06-scFv (n=12) mice).

    Techniques Used: Mouse Assay, Staining, One-tailed Test

    Plasma E06-scFv binds to atherosclerotic lesions and apoptotic thymocytes and is present in aorta of Ldlr −/− /E06-scFv mice a , Staining of atherosclerotic lesions of WHHL rabbit aorta with E06-scFv plasma (left panel), and Ldlr −/− mice (right panel) (both at dilution of 1:20), visualized using biotinylated anti-Myc mAb and ABC-AP VectaStain kit. b , Deconvolution microscopy of E06-scFv plasma (1:20 dilution) binding to apoptotic but not normal cells. Blue, nuclei stained with Hoechst dye; Green, FITC-labeled anti-His tag mAb; Red, Annexin V-PE. c , Binding of E06-scFv plasma (1:20 dilution) to apoptotic thymocytes (7AAD+/Annexin V+) by FACS analysis. d , Expression of E06-scFv in aortic lesion of Ldlr −/− / E06-scFv but not Ldlr −/− mouse. Cross-sections at the AV were stained with biotinylated anti-Myc mAb to identify presence of E06-scFv in atherosclerotic lesion. Nuclei counterstained using Hematoxylin QS (Original ×200). Panels a-c are representative of similar studies with 5 other plasma samples. Panel d is representative of studies in 3 other aortic sections.
    Figure Legend Snippet: Plasma E06-scFv binds to atherosclerotic lesions and apoptotic thymocytes and is present in aorta of Ldlr −/− /E06-scFv mice a , Staining of atherosclerotic lesions of WHHL rabbit aorta with E06-scFv plasma (left panel), and Ldlr −/− mice (right panel) (both at dilution of 1:20), visualized using biotinylated anti-Myc mAb and ABC-AP VectaStain kit. b , Deconvolution microscopy of E06-scFv plasma (1:20 dilution) binding to apoptotic but not normal cells. Blue, nuclei stained with Hoechst dye; Green, FITC-labeled anti-His tag mAb; Red, Annexin V-PE. c , Binding of E06-scFv plasma (1:20 dilution) to apoptotic thymocytes (7AAD+/Annexin V+) by FACS analysis. d , Expression of E06-scFv in aortic lesion of Ldlr −/− / E06-scFv but not Ldlr −/− mouse. Cross-sections at the AV were stained with biotinylated anti-Myc mAb to identify presence of E06-scFv in atherosclerotic lesion. Nuclei counterstained using Hematoxylin QS (Original ×200). Panels a-c are representative of similar studies with 5 other plasma samples. Panel d is representative of studies in 3 other aortic sections.

    Techniques Used: Mouse Assay, Staining, Microscopy, Binding Assay, Labeling, FACS, Expressing

    35) Product Images from "The Heat Shock Response and Chaperones/Heat Shock Proteins in Brain Tumors: Surface Expression, Release, and Possible Immune Consequences"

    Article Title: The Heat Shock Response and Chaperones/Heat Shock Proteins in Brain Tumors: Surface Expression, Release, and Possible Immune Consequences

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3588-07.2007

    Cell surface biotinylation reveals that GRP78, HSPs 27 and 70, and HspBP1 are on the surfaces of brain tumor cell lines. A , Using membrane-impermeant biotin, the cell lines shown were surface biotinylated, and surface proteins were enriched on streptavidin matrices. Biotinylated proteins were separated on gels and Western blotted. Antibody probes are shown at left. B , To verify that only surface proteins were labeled, the fractions unbound on the streptavidin beads (D54MG lysate) and the bound/eluted fractions (D54MG biotin) were probed with antibodies against cytosolic proteins tubulin, actin, and glyceraldehyde-3 phosphate dehydrogenase (GAPDH). HSP/HSC70 was found in both the nonbiotinylated cytosolic fraction and the labeled membrane fraction (as expected, from the results in A ).
    Figure Legend Snippet: Cell surface biotinylation reveals that GRP78, HSPs 27 and 70, and HspBP1 are on the surfaces of brain tumor cell lines. A , Using membrane-impermeant biotin, the cell lines shown were surface biotinylated, and surface proteins were enriched on streptavidin matrices. Biotinylated proteins were separated on gels and Western blotted. Antibody probes are shown at left. B , To verify that only surface proteins were labeled, the fractions unbound on the streptavidin beads (D54MG lysate) and the bound/eluted fractions (D54MG biotin) were probed with antibodies against cytosolic proteins tubulin, actin, and glyceraldehyde-3 phosphate dehydrogenase (GAPDH). HSP/HSC70 was found in both the nonbiotinylated cytosolic fraction and the labeled membrane fraction (as expected, from the results in A ).

    Techniques Used: Western Blot, Labeling

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    Avidin-Biotin Assay:

    Article Title: Surface Plasmon Resonance (SPR) Detection Using Antibody-Linked Magnetic Nanoparticles for Analyte Capture, Purification, Concentration and Signal Amplification
    Article Snippet: .. The unbiotinylated monoclonal anti-SEB antibodies served as a control and exhibited no avidin binding, while the commercially biotinylated antibodies from Vector Labs and the biotinylated monoclonal anti-SEB each bound similar amounts of avidin. .. Biotinylated antibodies were then mixed with 50 nanometer streptavidin-coated nanobeads (Miltenyi μMACS, Gladbach, Germany #120-001-017) by incubating 100 #l of the bead solution with an excess (50 μg) of biotinylated antibodies.

    Blocking Assay:

    Article Title: Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice
    Article Snippet: .. The blocked tissue was then incubated overnight with primary antibodies, washed with PBS, and exposed with intervening washes in PBS to the biotinylated secondary antibodies (2 h), 0.3% H2 O2 in 0.3% blocking sera (5 min), Elite ABC reagent (1 h; Vector Laboratories), and 3′,5′-diaminobenzidine solution (2–10 min) until suitable staining developed. .. As a control for the specificity of antibodies to the D2 and D3 receptors, attempts were made to immunostain the corresponding receptors in the gut of knock-out animals lacking these receptors.

    Concentration Assay:

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    Article Snippet: .. Following fixation, retinas were incubated in 0.1% Triton X-100 in PBS (0.1 M, pH 7.4) containing biotinylated anti-rhodamine (C# BA-0605, Vector Labs) at a concentration of 1:100 for 36 hr at room temperature, then washed in phosphate buffer overnight. .. Retinas were then processed for HRP histochemistry as described above using the Vector avidin-biotin-HRP complex and DAB.

    Incubation:

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    Article Snippet: .. Following antigen retrieval where necessary, sections were blocked with 5% normal serum, and incubated with the primary antibody for 1 hour, followed by biotinylated secondary antibody (Vector Laboratories, Burlingame, CA) diluted 1:500, and streptavidin HRP (Vector Laboratories) diluted 1:50. .. Sections were developed in Nova Red peroxidase substrate (Vector Laboratories) and counterstained with hematoxylin.

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    Staining:

    Article Title: Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice
    Article Snippet: .. The blocked tissue was then incubated overnight with primary antibodies, washed with PBS, and exposed with intervening washes in PBS to the biotinylated secondary antibodies (2 h), 0.3% H2 O2 in 0.3% blocking sera (5 min), Elite ABC reagent (1 h; Vector Laboratories), and 3′,5′-diaminobenzidine solution (2–10 min) until suitable staining developed. .. As a control for the specificity of antibodies to the D2 and D3 receptors, attempts were made to immunostain the corresponding receptors in the gut of knock-out animals lacking these receptors.

    Binding Assay:

    Article Title: Age-related increases in ozone-induced injury and altered pulmonary mechanics in mice with progressive lung inflammation
    Article Snippet: .. Sections were then incubated with biotinylated secondary antibody (1:200, Vector Labs, Burlingame, CA) for 30 min. A DAB Peroxidase Substrate Kit (Vector Labs) was used to visualize binding. ..

    Article Title: Surface Plasmon Resonance (SPR) Detection Using Antibody-Linked Magnetic Nanoparticles for Analyte Capture, Purification, Concentration and Signal Amplification
    Article Snippet: .. The unbiotinylated monoclonal anti-SEB antibodies served as a control and exhibited no avidin binding, while the commercially biotinylated antibodies from Vector Labs and the biotinylated monoclonal anti-SEB each bound similar amounts of avidin. .. Biotinylated antibodies were then mixed with 50 nanometer streptavidin-coated nanobeads (Miltenyi μMACS, Gladbach, Germany #120-001-017) by incubating 100 #l of the bead solution with an excess (50 μg) of biotinylated antibodies.

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  • 99
    Vector Laboratories biotinylated secondary antibodies
    D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a <t>biotinylated</t> donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.
    Biotinylated Secondary Antibodies, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated secondary antibodies/product/Vector Laboratories
    Average 99 stars, based on 1500 article reviews
    Price from $9.99 to $1999.99
    biotinylated secondary antibodies - by Bioz Stars, 2020-07
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    99
    Vector Laboratories vectashield hardset mounting medium
    Representative examples of immunocytochemistry for NSE, GFAP and 2B3 in mouse primary cortical cultures (A-D). (A) Specific labelling of a large population of wild-type neurones with an anti-NSE antibody (10μg/ml) detected using a biotinylated anti-rabbit antibody (1:500) and Avidin-FITC (1:600); (B) specific labelling of a small population of wild-type glial cells with an anti-GFAP antibody (1μg/ml) detected as above; (C) specific labelling of wild-type cells with 2B3 (10μg/ml) detected using a biotinylated anti-mouse antibody (1:500) and Avidin-FITC (1:600); (D) negative control for 2B3 showing low levels of non-specific staining in wild-type cells incubated in the absence of the primary antibody and detected as above. Similar results were obtained with negative controls for NSE and GFAP (data not shown). Coverslips were mounted with <t>VECTASHIELD</t> mounting medium with DAPI to counter-stain the nuclei. n = 5-7. Scale bar = 25μm.
    Vectashield Hardset Mounting Medium, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vectashield hardset mounting medium/product/Vector Laboratories
    Average 99 stars, based on 220 article reviews
    Price from $9.99 to $1999.99
    vectashield hardset mounting medium - by Bioz Stars, 2020-07
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    Image Search Results


    D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a biotinylated donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.

    Journal: The Journal of Neuroscience

    Article Title: Physiological Modulation of Intestinal Motility by Enteric Dopaminergic Neurons and the D2 Receptor: Analysis of Dopamine Receptor Expression, Location, Development, and Function in Wild-Type and Knock-Out Mice

    doi: 10.1523/JNEUROSCI.4720-05.2006

    Figure Lengend Snippet: D 2 and D 3 immunocytochemistry were performed on the submucosal plexus of the ileum of CD-1 mice. The same tissue preparation from D 2 and D 3 KO mice was used as control. D 2 receptor immunoreactivity (IR) was revealed by a D 2 rabbit antibody and a donkey anti-rabbit Alexa 594 secondary antibody. D 3 receptor immunoreactivity was revealed by a D 3 goat antibody, a biotinylated donkey anti-goat secondary antibody, and streptavidin FITC. On the tissue of CD-1 mice, D 2 -immunoreactive products were present in the enteric neurons ( A ), and D 3 -immunoreactive products were also present in the enteric neurons ( B ). D 2 and D 3 immunoreactivities were colocalized in the same cell ( C ). However, D 2 immunoreactivity was not detected on the tissue of D 2 knock-out (KO) mice ( D ); no D 3 immunoreactivity was detected on the tissue of D 3 knock-out mouse ( E ). The arrows indicate the immunoreactive neurons. Scale bar: (in C ) 25 μm.

    Article Snippet: The blocked tissue was then incubated overnight with primary antibodies, washed with PBS, and exposed with intervening washes in PBS to the biotinylated secondary antibodies (2 h), 0.3% H2 O2 in 0.3% blocking sera (5 min), Elite ABC reagent (1 h; Vector Laboratories), and 3′,5′-diaminobenzidine solution (2–10 min) until suitable staining developed.

    Techniques: Immunocytochemistry, Mouse Assay, Knock-Out

    Immunohistochemical staining of human MPM tissue using the avidin-biotin-peroxidase complex procedure. (A) A Wnt1-high tumor. A tumor with positive expression of (B) Wnt2B, (C) nuclear survivin and (D) c-Myc. A tumor with negative expression of (E) Wnt2B, (F) nuclear survivin and (G) c-Myc. (H) A Wnt5A-high tumor. Original magnification, ×200.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Intratumoral Wnt2B expression affects tumor proliferation and survival in malignant pleural mesothelioma patients

    doi: 10.3892/etm.2012.511

    Figure Lengend Snippet: Immunohistochemical staining of human MPM tissue using the avidin-biotin-peroxidase complex procedure. (A) A Wnt1-high tumor. A tumor with positive expression of (B) Wnt2B, (C) nuclear survivin and (D) c-Myc. A tumor with negative expression of (E) Wnt2B, (F) nuclear survivin and (G) c-Myc. (H) A Wnt5A-high tumor. Original magnification, ×200.

    Article Snippet: Slides were then incubated for 1 h with biotinylated secondary antibodies (Vector Laboratories, Burlingame, CA, USA).

    Techniques: Immunohistochemistry, Staining, Avidin-Biotin Assay, Expressing

    Representative examples of immunocytochemistry for NSE, GFAP and 2B3 in mouse primary cortical cultures (A-D). (A) Specific labelling of a large population of wild-type neurones with an anti-NSE antibody (10μg/ml) detected using a biotinylated anti-rabbit antibody (1:500) and Avidin-FITC (1:600); (B) specific labelling of a small population of wild-type glial cells with an anti-GFAP antibody (1μg/ml) detected as above; (C) specific labelling of wild-type cells with 2B3 (10μg/ml) detected using a biotinylated anti-mouse antibody (1:500) and Avidin-FITC (1:600); (D) negative control for 2B3 showing low levels of non-specific staining in wild-type cells incubated in the absence of the primary antibody and detected as above. Similar results were obtained with negative controls for NSE and GFAP (data not shown). Coverslips were mounted with VECTASHIELD mounting medium with DAPI to counter-stain the nuclei. n = 5-7. Scale bar = 25μm.

    Journal: Neuroreport

    Article Title: Inhibition of amyloid-β production by anti-amyloid precursor protein antibodies in primary mouse cortical neurones

    doi: 10.1097/WNR.0000000000000055

    Figure Lengend Snippet: Representative examples of immunocytochemistry for NSE, GFAP and 2B3 in mouse primary cortical cultures (A-D). (A) Specific labelling of a large population of wild-type neurones with an anti-NSE antibody (10μg/ml) detected using a biotinylated anti-rabbit antibody (1:500) and Avidin-FITC (1:600); (B) specific labelling of a small population of wild-type glial cells with an anti-GFAP antibody (1μg/ml) detected as above; (C) specific labelling of wild-type cells with 2B3 (10μg/ml) detected using a biotinylated anti-mouse antibody (1:500) and Avidin-FITC (1:600); (D) negative control for 2B3 showing low levels of non-specific staining in wild-type cells incubated in the absence of the primary antibody and detected as above. Similar results were obtained with negative controls for NSE and GFAP (data not shown). Coverslips were mounted with VECTASHIELD mounting medium with DAPI to counter-stain the nuclei. n = 5-7. Scale bar = 25μm.

    Article Snippet: Cover slips were mounted on to glass slides using VECTASHIELD® HardSet™ Mounting Medium (Vector Laboratories) with DAPI (1:4) and visualized using a Leica DMIRE2 microscope and Openlab 4.04 software.

    Techniques: Immunocytochemistry, Avidin-Biotin Assay, Negative Control, Staining, Incubation

    Compatibility of Polymer Dye Labeling with Vectashield mounting medium. Polymer Dye Labeling of nuclear pore complex imaged (a) dry or (b) in Vectashield hardset mounting medium. Polymer coated nuclei without Evans Blue dye imaged (c) dry or (d) in Vectashield hardset mounting medium. Scale bars are 50 µm.

    Journal: PLoS ONE

    Article Title: Interfacial Polymerization for Colorimetric Labeling of Protein Expression in Cells

    doi: 10.1371/journal.pone.0115630

    Figure Lengend Snippet: Compatibility of Polymer Dye Labeling with Vectashield mounting medium. Polymer Dye Labeling of nuclear pore complex imaged (a) dry or (b) in Vectashield hardset mounting medium. Polymer coated nuclei without Evans Blue dye imaged (c) dry or (d) in Vectashield hardset mounting medium. Scale bars are 50 µm.

    Article Snippet: Biotinylated polyclonal goat IgG anti-mouse IgG (H+L; catalogue #BA-9200) and Vectashield hardset mounting medium was purchased from Vector Laboratories (Burlingame, CA).

    Techniques: Labeling