goat anti rabbit igg  (Thermo Fisher)


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    Structured Review

    Thermo Fisher goat anti rabbit igg
    The number fraction of <t>rAb-MP</t> aggregates to all particles as a function of goat <t>IgG</t> concentration in PBS containing 0.1% BSA. Particle counts were obtained from bright field microscope images. The standard deviation was calculated from three replicates.
    Goat Anti Rabbit Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 239 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg/product/Thermo Fisher
    Average 99 stars, based on 239 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg - by Bioz Stars, 2020-02
    99/100 stars

    Images

    1) Product Images from "A Versatile Microparticle-Based Immunoaggregation Assay for Macromolecular Biomarker Detection and Quantification"

    Article Title: A Versatile Microparticle-Based Immunoaggregation Assay for Macromolecular Biomarker Detection and Quantification

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0115046

    The number fraction of rAb-MP aggregates to all particles as a function of goat IgG concentration in PBS containing 0.1% BSA. Particle counts were obtained from bright field microscope images. The standard deviation was calculated from three replicates.
    Figure Legend Snippet: The number fraction of rAb-MP aggregates to all particles as a function of goat IgG concentration in PBS containing 0.1% BSA. Particle counts were obtained from bright field microscope images. The standard deviation was calculated from three replicates.

    Techniques Used: Concentration Assay, Microscopy, Standard Deviation

    Accusizer measurement results for (A) FITC labeled rAb-MP without goat Ig G and (B) FITC labeled rAb-MP with 36 ng/mL goat IgG as a model macromolecular biomarker. The concentration of rAb-MP was kept constant at 53.4 μg/mL.
    Figure Legend Snippet: Accusizer measurement results for (A) FITC labeled rAb-MP without goat Ig G and (B) FITC labeled rAb-MP with 36 ng/mL goat IgG as a model macromolecular biomarker. The concentration of rAb-MP was kept constant at 53.4 μg/mL.

    Techniques Used: Labeling, Biomarker Assay, Concentration Assay

    Fluorescence Microscope images: (A) FITC labeled rAb-MP without goat Ig G and (B) FITC labeled rAb-MP with 36 ng/mL goat IgG as a model biomarker. The concentration of rAb-MP was kept constant at 53.4 μg/mL.
    Figure Legend Snippet: Fluorescence Microscope images: (A) FITC labeled rAb-MP without goat Ig G and (B) FITC labeled rAb-MP with 36 ng/mL goat IgG as a model biomarker. The concentration of rAb-MP was kept constant at 53.4 μg/mL.

    Techniques Used: Fluorescence, Microscopy, Labeling, Biomarker Assay, Concentration Assay

    2) Product Images from "Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG"

    Article Title: Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00297-13

    Evaluation of the ability of purified IgM and IgG from immune sera to inhibit C. burnetii infection in vivo by comparing splenomegaly as well as bacterial burden and pathological changes in the spleen with those of normal mouse IgM and IgG controls at
    Figure Legend Snippet: Evaluation of the ability of purified IgM and IgG from immune sera to inhibit C. burnetii infection in vivo by comparing splenomegaly as well as bacterial burden and pathological changes in the spleen with those of normal mouse IgM and IgG controls at

    Techniques Used: Purification, Infection, In Vivo

    3) Product Images from "Fibroblast Growth Factor Homologous Factor 2B: Association with Nav1.6 and Selective Colocalization at Nodes of Ranvier of Dorsal Root Axons"

    Article Title: Fibroblast Growth Factor Homologous Factor 2B: Association with Nav1.6 and Selective Colocalization at Nodes of Ranvier of Dorsal Root Axons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1628-04.2004

    FHF2 and Na v 1.6 are colocalized in the brain. A , Pan sodium channel and FHF2 antibodies were used to immunoprecipitate (IP) the voltage-gated sodium channels from a rat brain lysate. Anti-mouse IgG was used as a negative control to rule out nonspecific binding. Western blotting analysis of the IP complex was performed by the pan sodium channel antibody. Lane 1 shows a robust immunoreactive signal from the cell lysate that was used for the IP assay, consistent with the presence of intact sodium channel proteins in this sample. Nonspecific antibodies do not immunoprecipitate a channel complex (lane 2). As predicted, pan sodium channel antibody immunoprecipitated a channel complex (lane 3). The reduced signal in lane 3 compared with lane 1 is likely attributable to the avidity of the antibody in the two different assays. FHF2 coimmunoprecipitated voltage-gated sodium channels from the brain lysate (lane 4). Comparison of the immunoreactive bands in lanes 1 and 4 might be explained by the interaction of FHF2 with only a subset of the cellular pool of channels. The molecular weight marker (in kilodaltons) is shown on the left. B , Colocalization of Na v 1.6 and FHF2 in the hippocampus. Na v 1.6- and FHF2-specific antibodies were used to immunolabel sections of rat hippocampus. FHF2 (green) and Na v 1.6 (red) proteins were detected in the pyramidal cells of Ammon's horn, and the merged images (yellow) show significant colocalization of the two proteins. The inset shows the pyramidal cells at 40× magnification. C , GFP antibody was used to IP Na v 1.6 from lysates of HEK 293 cells transfected with either Na v 1.6 plus GFP (control; lane 3) or Na v 1.6 plus FHF2B-GFP (lane 4). The IP complex was probed with the pan sodium channel antibody and detected no association between Na v 1.6 with GFP (lane 3), but an association of Na v 1.6 with FHF2B (lane 4). Lanes 1 and 2 show Western blotting analysis of the cell lysates probed with pan sodium channel antibody (top) and GFP (bottom) to show comparable levels of Na v 1.6/GFP (lane 1) and Na v 1.6/FHF2B-GFP (lane 2) in the samples used for the immunoprecipitation assay.
    Figure Legend Snippet: FHF2 and Na v 1.6 are colocalized in the brain. A , Pan sodium channel and FHF2 antibodies were used to immunoprecipitate (IP) the voltage-gated sodium channels from a rat brain lysate. Anti-mouse IgG was used as a negative control to rule out nonspecific binding. Western blotting analysis of the IP complex was performed by the pan sodium channel antibody. Lane 1 shows a robust immunoreactive signal from the cell lysate that was used for the IP assay, consistent with the presence of intact sodium channel proteins in this sample. Nonspecific antibodies do not immunoprecipitate a channel complex (lane 2). As predicted, pan sodium channel antibody immunoprecipitated a channel complex (lane 3). The reduced signal in lane 3 compared with lane 1 is likely attributable to the avidity of the antibody in the two different assays. FHF2 coimmunoprecipitated voltage-gated sodium channels from the brain lysate (lane 4). Comparison of the immunoreactive bands in lanes 1 and 4 might be explained by the interaction of FHF2 with only a subset of the cellular pool of channels. The molecular weight marker (in kilodaltons) is shown on the left. B , Colocalization of Na v 1.6 and FHF2 in the hippocampus. Na v 1.6- and FHF2-specific antibodies were used to immunolabel sections of rat hippocampus. FHF2 (green) and Na v 1.6 (red) proteins were detected in the pyramidal cells of Ammon's horn, and the merged images (yellow) show significant colocalization of the two proteins. The inset shows the pyramidal cells at 40× magnification. C , GFP antibody was used to IP Na v 1.6 from lysates of HEK 293 cells transfected with either Na v 1.6 plus GFP (control; lane 3) or Na v 1.6 plus FHF2B-GFP (lane 4). The IP complex was probed with the pan sodium channel antibody and detected no association between Na v 1.6 with GFP (lane 3), but an association of Na v 1.6 with FHF2B (lane 4). Lanes 1 and 2 show Western blotting analysis of the cell lysates probed with pan sodium channel antibody (top) and GFP (bottom) to show comparable levels of Na v 1.6/GFP (lane 1) and Na v 1.6/FHF2B-GFP (lane 2) in the samples used for the immunoprecipitation assay.

    Techniques Used: Negative Control, Binding Assay, Western Blot, Immunoprecipitation, Molecular Weight, Marker, Immunolabeling, Transfection

    4) Product Images from "DC3, the 21-kDa Subunit of the Outer Dynein Arm-Docking Complex (ODA-DC), Is a Novel EF-Hand Protein Important for Assembly of Both the Outer Arm and the ODA-DC"

    Article Title: DC3, the 21-kDa Subunit of the Outer Dynein Arm-Docking Complex (ODA-DC), Is a Novel EF-Hand Protein Important for Assembly of Both the Outer Arm and the ODA-DC

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E03-01-0057

    (A) DC1 and DC2 assemble on the axoneme in the absence of DC3, but not vice versa. Top panel: Axonemes from wild type, the DC3-deletion strain (DC3Δ), oda 1, and oda 3 were isolated and analyzed by western blotting. oda 1 is null for DC2; oda 3 is null for DC1. The blot was probed with antibodies to DC1, DC2, and the inner arm IC, IC140 (used as a loading control). As expected, antibodies to DC1 detected protein in wild-type axonemes, but not in oda 1 or oda 3 axonemes, which lack the ODA-DC. Importantly, the antibody also detected protein in axonemes of the DC3-deletion strain. Essentially identical results were obtained with antibodies to DC2 (our unpublished results). These data indicate that DC1 and DC2 can assemble on the axoneme in the complete absence of DC3. Bottom panel: Axonemes from wild type, the DC3-deletion strain (DC3Δ), oda 1, oda 3, and oda 9 were prepared as above ( oda 9 has a defect in IC1 and lacks outer dynein arms but retains the ODA-DC). The blot was probed with a polyclonal antibody to DC3. The antibody detects a single protein of M r ∼25,000 in both oda 9 and wild-type axonemes but detects no protein in DC3-null axonemes, indicating that it is specific for DC3. Axonemes from oda 1 and oda 3 do not contain DC3, indicating that DC3 assembly is dependent on the presence of DC1 and DC2. (B) Immunoprecipitation of the ODA-DC in the absence of Mg 2 + . The DC1 antibody was used to immunoprecipitate the ODA-DC from biotinylated 0.6 M KCl extracts of DC3-transformant (strain W215) and DC3-null axonemes. The immunoprecipitated proteins were then resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and detected using streptavidin-HRP. The anti-DC1 antibody immuno-precipitated three proteins of M r ∼105,000, ∼70,000, and ∼25,000 (arrowheads) from DC3-transformant axonemal extracts (lane 3). These proteins, corresponding to DC1, DC2, and DC3, respectively, were not immunoprecipitated from DC3-transformant axonemal extracts using normal rabbit IgG (lane 4). In contrast, the anti-DC1 antibody immunoprecipitated only DC1 and DC2 (arrowheads) from the DC3-null axonemal extracts (lane 1). These proteins were not immunoprecipitated from DC3-null axonemal extracts using normal rabbit IgG (lane 2). These data confirm that a “partial” docking complex composed of DC1 and DC2 assembles on the axoneme when DC3 is missing and that transformation of the DC3-deletion strain with the DC3 gene restores DC3 to the ODA-DC. Numbers on left indicate molecular weight markers.
    Figure Legend Snippet: (A) DC1 and DC2 assemble on the axoneme in the absence of DC3, but not vice versa. Top panel: Axonemes from wild type, the DC3-deletion strain (DC3Δ), oda 1, and oda 3 were isolated and analyzed by western blotting. oda 1 is null for DC2; oda 3 is null for DC1. The blot was probed with antibodies to DC1, DC2, and the inner arm IC, IC140 (used as a loading control). As expected, antibodies to DC1 detected protein in wild-type axonemes, but not in oda 1 or oda 3 axonemes, which lack the ODA-DC. Importantly, the antibody also detected protein in axonemes of the DC3-deletion strain. Essentially identical results were obtained with antibodies to DC2 (our unpublished results). These data indicate that DC1 and DC2 can assemble on the axoneme in the complete absence of DC3. Bottom panel: Axonemes from wild type, the DC3-deletion strain (DC3Δ), oda 1, oda 3, and oda 9 were prepared as above ( oda 9 has a defect in IC1 and lacks outer dynein arms but retains the ODA-DC). The blot was probed with a polyclonal antibody to DC3. The antibody detects a single protein of M r ∼25,000 in both oda 9 and wild-type axonemes but detects no protein in DC3-null axonemes, indicating that it is specific for DC3. Axonemes from oda 1 and oda 3 do not contain DC3, indicating that DC3 assembly is dependent on the presence of DC1 and DC2. (B) Immunoprecipitation of the ODA-DC in the absence of Mg 2 + . The DC1 antibody was used to immunoprecipitate the ODA-DC from biotinylated 0.6 M KCl extracts of DC3-transformant (strain W215) and DC3-null axonemes. The immunoprecipitated proteins were then resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and detected using streptavidin-HRP. The anti-DC1 antibody immuno-precipitated three proteins of M r ∼105,000, ∼70,000, and ∼25,000 (arrowheads) from DC3-transformant axonemal extracts (lane 3). These proteins, corresponding to DC1, DC2, and DC3, respectively, were not immunoprecipitated from DC3-transformant axonemal extracts using normal rabbit IgG (lane 4). In contrast, the anti-DC1 antibody immunoprecipitated only DC1 and DC2 (arrowheads) from the DC3-null axonemal extracts (lane 1). These proteins were not immunoprecipitated from DC3-null axonemal extracts using normal rabbit IgG (lane 2). These data confirm that a “partial” docking complex composed of DC1 and DC2 assembles on the axoneme when DC3 is missing and that transformation of the DC3-deletion strain with the DC3 gene restores DC3 to the ODA-DC. Numbers on left indicate molecular weight markers.

    Techniques Used: Isolation, Western Blot, Immunoprecipitation, SDS Page, Transformation Assay, Molecular Weight

    5) Product Images from "The extradomain a of fibronectin enhances the efficacy of lipopolysaccharide defective Salmonella bacterins as vaccines in mice"

    Article Title: The extradomain a of fibronectin enhances the efficacy of lipopolysaccharide defective Salmonella bacterins as vaccines in mice

    Journal: Veterinary Research

    doi: 10.1186/1297-9716-43-31

    EDAvidin binding to biotinylated bacterins in ELISA. ELISA plates coated with biotinylated (Biot) or not biotinylated control (B-SEΔ waaL , B-SEΔ gal and B-SEwt) bacterins were incubated with EDAvidin or EDA alone (control). Binding was monitored using a rabbit anti-EDA polyclonal antibody and an anti-rabbit whole IgG horseradish-peroxidase-conjugated second antibody. The O.D. values at 405 nm (mean ± SD) are represented.
    Figure Legend Snippet: EDAvidin binding to biotinylated bacterins in ELISA. ELISA plates coated with biotinylated (Biot) or not biotinylated control (B-SEΔ waaL , B-SEΔ gal and B-SEwt) bacterins were incubated with EDAvidin or EDA alone (control). Binding was monitored using a rabbit anti-EDA polyclonal antibody and an anti-rabbit whole IgG horseradish-peroxidase-conjugated second antibody. The O.D. values at 405 nm (mean ± SD) are represented.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

    6) Product Images from "IMMUNODOMINANT EPITOPE AND PROPERTIES OF PYROGLUTAMATE-MODIFIED A?-SPECIFIC ANTIBODIES PRODUCED IN RABBITS"

    Article Title: IMMUNODOMINANT EPITOPE AND PROPERTIES OF PYROGLUTAMATE-MODIFIED A?-SPECIFIC ANTIBODIES PRODUCED IN RABBITS

    Journal: Journal of neuroimmunology

    doi: 10.1016/j.jneuroim.2009.06.003

    Rabbit anti-AβN3(pE) IgG are specifically binding in ELISA to AβN3(pE) peptide, while recognition of AβN11(pE) and Aβ 1-42 is negligible. AβN3(pE), AβN11(pE) and Aβ 1-42 were prepared as described in Materials and methods and used for covering microtiter plates. Optical densities (OD) were registered at 405. Data are means of three independent experiments ±SD.
    Figure Legend Snippet: Rabbit anti-AβN3(pE) IgG are specifically binding in ELISA to AβN3(pE) peptide, while recognition of AβN11(pE) and Aβ 1-42 is negligible. AβN3(pE), AβN11(pE) and Aβ 1-42 were prepared as described in Materials and methods and used for covering microtiter plates. Optical densities (OD) were registered at 405. Data are means of three independent experiments ±SD.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay

    7) Product Images from "Knockdown of lncRNA-ATB suppresses autocrine secretion of TGF-β2 by targeting ZNF217 via miR-200c in keloid fibroblasts"

    Article Title: Knockdown of lncRNA-ATB suppresses autocrine secretion of TGF-β2 by targeting ZNF217 via miR-200c in keloid fibroblasts

    Journal: Scientific Reports

    doi: 10.1038/srep24728

    ZNF217 is a transcriptional activator of TGF-β2 in keloid fibroblasts. ( A ) Schematic representation of ZNF217 binding site in the TGF-β2 promoter region. TSS: transcription start site; ( B ) ChIP assays using anti-ZNF217 antibody or human IgG as a control on the TGF-β2 promoter region, followed by PCR using ZNF217 binding site-specific primers for measurement of TGF-β2 expression; ( C ) qRT-PCR determination of relative ZNF217 expression levels in keloid fibroblasts (KF) treated with a control siRNA (siRNA-control) or ZNF217 specific siRNAs (siRNA-ZNF217-1 and siRNA- ZNF217-2); ( D , E ) qRT-PCR and ELISA determination of relative TGF-β2 mRNA and protein expression levels in KF treated with ZNF217-specific siRNAs, respectively.
    Figure Legend Snippet: ZNF217 is a transcriptional activator of TGF-β2 in keloid fibroblasts. ( A ) Schematic representation of ZNF217 binding site in the TGF-β2 promoter region. TSS: transcription start site; ( B ) ChIP assays using anti-ZNF217 antibody or human IgG as a control on the TGF-β2 promoter region, followed by PCR using ZNF217 binding site-specific primers for measurement of TGF-β2 expression; ( C ) qRT-PCR determination of relative ZNF217 expression levels in keloid fibroblasts (KF) treated with a control siRNA (siRNA-control) or ZNF217 specific siRNAs (siRNA-ZNF217-1 and siRNA- ZNF217-2); ( D , E ) qRT-PCR and ELISA determination of relative TGF-β2 mRNA and protein expression levels in KF treated with ZNF217-specific siRNAs, respectively.

    Techniques Used: Binding Assay, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    8) Product Images from "Epstein–Barr Virus Lytic Reactivation Induces IgG4 Production by Host B Lymphocytes in Graves' Disease Patients and Controls: A Subset of Graves' Disease Is an IgG4-Related Disease-Like Condition"

    Article Title: Epstein–Barr Virus Lytic Reactivation Induces IgG4 Production by Host B Lymphocytes in Graves' Disease Patients and Controls: A Subset of Graves' Disease Is an IgG4-Related Disease-Like Condition

    Journal: Viral Immunology

    doi: 10.1089/vim.2018.0042

    EBER1-positive cells and IgG4-positive cells were observed in the same areas. Resected tissues showed the diffuse hyperplasia of thyroid follicular epithelial cells with the focal infiltration of lymphocytes, but not tumefactive lesions, storiform fibrosis, or obliterative phlebitis (A) . EBER1-positive cells and IgG4-positive plasma cells were observed in the same area with lymphoid cell infiltration (B, D) . Six of the seven cases had a large number of IgG4-positive plasma cells (10/HPF
    Figure Legend Snippet: EBER1-positive cells and IgG4-positive cells were observed in the same areas. Resected tissues showed the diffuse hyperplasia of thyroid follicular epithelial cells with the focal infiltration of lymphocytes, but not tumefactive lesions, storiform fibrosis, or obliterative phlebitis (A) . EBER1-positive cells and IgG4-positive plasma cells were observed in the same area with lymphoid cell infiltration (B, D) . Six of the seven cases had a large number of IgG4-positive plasma cells (10/HPF

    Techniques Used:

    Production of IgG and IgG4 during the EBV reactivation period. Culture fluids were sampled on days 0, 5, 10, and 12, and IgG (A) and IgG4 (B) were then measured by ELISA. Time course changes were significant in Friedman's analysis of variance. IgG4 percentages (C) were higher than normal serum levels (approximately 4%). However, no significant difference was observed between patients and controls (D) .
    Figure Legend Snippet: Production of IgG and IgG4 during the EBV reactivation period. Culture fluids were sampled on days 0, 5, 10, and 12, and IgG (A) and IgG4 (B) were then measured by ELISA. Time course changes were significant in Friedman's analysis of variance. IgG4 percentages (C) were higher than normal serum levels (approximately 4%). However, no significant difference was observed between patients and controls (D) .

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Detection of IgG4(+)72A1(+) double-positive cells in culture cells on day 5. We detected IgG4-positive and EBV-reactivated [IgG4(+)72A1(+)] cells in culture cells on day 5 and confirmed sorted cells by confocal laser microscope. Red spots are surface IgG4 and fine green dots ).
    Figure Legend Snippet: Detection of IgG4(+)72A1(+) double-positive cells in culture cells on day 5. We detected IgG4-positive and EBV-reactivated [IgG4(+)72A1(+)] cells in culture cells on day 5 and confirmed sorted cells by confocal laser microscope. Red spots are surface IgG4 and fine green dots ).

    Techniques Used: Microscopy

    9) Product Images from "Advantages of Papio anubis for preclinical testing of immunotoxicity of candidate therapeutic antagonist antibodies targeting CD28"

    Article Title: Advantages of Papio anubis for preclinical testing of immunotoxicity of candidate therapeutic antagonist antibodies targeting CD28

    Journal: mAbs

    doi: 10.4161/mabs.28375

    Figure 6. In vitro response of human PBMC to antagonist anti-CD28 mAbs and ADA . Human PBMC were stimulated with anti-CD3 antibodies and different types of anti-CD28 mAbs plus indicated concentrations of IgG containing ADA. ( A ) Proliferation at
    Figure Legend Snippet: Figure 6. In vitro response of human PBMC to antagonist anti-CD28 mAbs and ADA . Human PBMC were stimulated with anti-CD3 antibodies and different types of anti-CD28 mAbs plus indicated concentrations of IgG containing ADA. ( A ) Proliferation at

    Techniques Used: In Vitro

    10) Product Images from "YB-1 regulates tumor growth by promoting MACC1/c-Met pathway in human lung adenocarcinoma"

    Article Title: YB-1 regulates tumor growth by promoting MACC1/c-Met pathway in human lung adenocarcinoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.18262

    YB-1 promotes MACC1 transcription by binding to MACC1 promoter and activates MACC1/c-Met pathway (A) Analysis of the MACC1 promoter indicated two putative YB-1 binding sites where the black boxes indicate sequences. RT-PCR analysis (B) and western blot analysis (C) determined the expression of YB-1 and MACC1 after inhibition of YB-1 in A549 cells. (D) MACC1 promoter (-2020 to +262) activity was analyzed by dual-luciferase reporter assay. The YB-1-silenced A549 cells and their corresponding control cells were co-transfected with plvx control plasmid or plvx-YB1 plasmid and MACC1 promoter (-2020 to +262) or basic reporter along with pRL-TK for 24h. (E) Mutated MACC1 promoter plasmids were generated (pGL3- MACC1 -mutated1; pGL3- MACC1 -mutated2). YB-1 binding sites in these two plasmids were mutated at two base pairs (pGL3- MACC1 -mutated1: -1860 to -1856; pGL3- MACC1 -mutated2: -1468 to -1464). A549 cells were transiently co-transfected with plvx vector plasmid or plvx-YB1 plasmid and the pGL3-basic reporter, the wild type MACC1 promoter reporter (pGL3- MACC1 ) or the mutated MACC1 promoter plasmids (pGL3-MACC1-mutated1; pGL3-MACC1-mutated2) along with pRL-TK for 24h. Transfected cells were harvested for dual-luciferase reporter assay. (F) ChIP assay was performed with YB-1 antibody or non-immune IgG as negative control. Immunoprecipitated DNA was amplified by PCR using primers as indicated. (G) Western blot analysis of YB-1, MACC1 and c-Met expression after inhibition of YB-1 in A549 cell. * P
    Figure Legend Snippet: YB-1 promotes MACC1 transcription by binding to MACC1 promoter and activates MACC1/c-Met pathway (A) Analysis of the MACC1 promoter indicated two putative YB-1 binding sites where the black boxes indicate sequences. RT-PCR analysis (B) and western blot analysis (C) determined the expression of YB-1 and MACC1 after inhibition of YB-1 in A549 cells. (D) MACC1 promoter (-2020 to +262) activity was analyzed by dual-luciferase reporter assay. The YB-1-silenced A549 cells and their corresponding control cells were co-transfected with plvx control plasmid or plvx-YB1 plasmid and MACC1 promoter (-2020 to +262) or basic reporter along with pRL-TK for 24h. (E) Mutated MACC1 promoter plasmids were generated (pGL3- MACC1 -mutated1; pGL3- MACC1 -mutated2). YB-1 binding sites in these two plasmids were mutated at two base pairs (pGL3- MACC1 -mutated1: -1860 to -1856; pGL3- MACC1 -mutated2: -1468 to -1464). A549 cells were transiently co-transfected with plvx vector plasmid or plvx-YB1 plasmid and the pGL3-basic reporter, the wild type MACC1 promoter reporter (pGL3- MACC1 ) or the mutated MACC1 promoter plasmids (pGL3-MACC1-mutated1; pGL3-MACC1-mutated2) along with pRL-TK for 24h. Transfected cells were harvested for dual-luciferase reporter assay. (F) ChIP assay was performed with YB-1 antibody or non-immune IgG as negative control. Immunoprecipitated DNA was amplified by PCR using primers as indicated. (G) Western blot analysis of YB-1, MACC1 and c-Met expression after inhibition of YB-1 in A549 cell. * P

    Techniques Used: Binding Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Inhibition, Activity Assay, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Generated, Chromatin Immunoprecipitation, Negative Control, Immunoprecipitation, Amplification, Polymerase Chain Reaction

    11) Product Images from "Defects in lysosomal maturation facilitate the activation of innate sensors in systemic lupus erythematosus"

    Article Title: Defects in lysosomal maturation facilitate the activation of innate sensors in systemic lupus erythematosus

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

    doi: 10.1073/pnas.1513943113

    On the surface of NOD myeloid cells, IgG and Sm show punctate staining. Splenic myeloid cells (CD11b + ) were purified from NOD mice and analyzed for surface levels of Sm and IgG by confocal microscopy; two experiments, two mice, 15–20 cells. Approximately
    Figure Legend Snippet: On the surface of NOD myeloid cells, IgG and Sm show punctate staining. Splenic myeloid cells (CD11b + ) were purified from NOD mice and analyzed for surface levels of Sm and IgG by confocal microscopy; two experiments, two mice, 15–20 cells. Approximately

    Techniques Used: Staining, Purification, Mouse Assay, Confocal Microscopy

    Autoimmune-prone MFs accumulate IgG-ICs on the cell membrane. ( A and B ) CD11b + cells were purified from spleen and analyzed for surface Sm and IgG by confocal imaging. (Scale bars: 2.5 μm.) Data represent two experiments, two mice, 8–10
    Figure Legend Snippet: Autoimmune-prone MFs accumulate IgG-ICs on the cell membrane. ( A and B ) CD11b + cells were purified from spleen and analyzed for surface Sm and IgG by confocal imaging. (Scale bars: 2.5 μm.) Data represent two experiments, two mice, 8–10

    Techniques Used: Purification, Imaging, Mouse Assay

    12) Product Images from "Fibroblast Growth Factor Homologous Factor 2B: Association with Nav1.6 and Selective Colocalization at Nodes of Ranvier of Dorsal Root Axons"

    Article Title: Fibroblast Growth Factor Homologous Factor 2B: Association with Nav1.6 and Selective Colocalization at Nodes of Ranvier of Dorsal Root Axons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1628-04.2004

    FHF2 and Na v 1.6 are colocalized in the brain. A , Pan sodium channel and FHF2 antibodies were used to immunoprecipitate (IP) the voltage-gated sodium channels from a rat brain lysate. Anti-mouse IgG was used as a negative control to rule out nonspecific binding. Western blotting analysis of the IP complex was performed by the pan sodium channel antibody. Lane 1 shows a robust immunoreactive signal from the cell lysate that was used for the IP assay, consistent with the presence of intact sodium channel proteins in this sample. Nonspecific antibodies do not immunoprecipitate a channel complex (lane 2). As predicted, pan sodium channel antibody immunoprecipitated a channel complex (lane 3). The reduced signal in lane 3 compared with lane 1 is likely attributable to the avidity of the antibody in the two different assays. FHF2 coimmunoprecipitated voltage-gated sodium channels from the brain lysate (lane 4). Comparison of the immunoreactive bands in lanes 1 and 4 might be explained by the interaction of FHF2 with only a subset of the cellular pool of channels. The molecular weight marker (in kilodaltons) is shown on the left. B , Colocalization of Na v 1.6 and FHF2 in the hippocampus. Na v 1.6- and FHF2-specific antibodies were used to immunolabel sections of rat hippocampus. FHF2 (green) and Na v 1.6 (red) proteins were detected in the pyramidal cells of Ammon's horn, and the merged images (yellow) show significant colocalization of the two proteins. The inset shows the pyramidal cells at 40× magnification. C , GFP antibody was used to IP Na v 1.6 from lysates of HEK 293 cells transfected with either Na v 1.6 plus GFP (control; lane 3) or Na v 1.6 plus FHF2B-GFP (lane 4). The IP complex was probed with the pan sodium channel antibody and detected no association between Na v 1.6 with GFP (lane 3), but an association of Na v 1.6 with FHF2B (lane 4). Lanes 1 and 2 show Western blotting analysis of the cell lysates probed with pan sodium channel antibody (top) and GFP (bottom) to show comparable levels of Na v 1.6/GFP (lane 1) and Na v 1.6/FHF2B-GFP (lane 2) in the samples used for the immunoprecipitation assay.
    Figure Legend Snippet: FHF2 and Na v 1.6 are colocalized in the brain. A , Pan sodium channel and FHF2 antibodies were used to immunoprecipitate (IP) the voltage-gated sodium channels from a rat brain lysate. Anti-mouse IgG was used as a negative control to rule out nonspecific binding. Western blotting analysis of the IP complex was performed by the pan sodium channel antibody. Lane 1 shows a robust immunoreactive signal from the cell lysate that was used for the IP assay, consistent with the presence of intact sodium channel proteins in this sample. Nonspecific antibodies do not immunoprecipitate a channel complex (lane 2). As predicted, pan sodium channel antibody immunoprecipitated a channel complex (lane 3). The reduced signal in lane 3 compared with lane 1 is likely attributable to the avidity of the antibody in the two different assays. FHF2 coimmunoprecipitated voltage-gated sodium channels from the brain lysate (lane 4). Comparison of the immunoreactive bands in lanes 1 and 4 might be explained by the interaction of FHF2 with only a subset of the cellular pool of channels. The molecular weight marker (in kilodaltons) is shown on the left. B , Colocalization of Na v 1.6 and FHF2 in the hippocampus. Na v 1.6- and FHF2-specific antibodies were used to immunolabel sections of rat hippocampus. FHF2 (green) and Na v 1.6 (red) proteins were detected in the pyramidal cells of Ammon's horn, and the merged images (yellow) show significant colocalization of the two proteins. The inset shows the pyramidal cells at 40× magnification. C , GFP antibody was used to IP Na v 1.6 from lysates of HEK 293 cells transfected with either Na v 1.6 plus GFP (control; lane 3) or Na v 1.6 plus FHF2B-GFP (lane 4). The IP complex was probed with the pan sodium channel antibody and detected no association between Na v 1.6 with GFP (lane 3), but an association of Na v 1.6 with FHF2B (lane 4). Lanes 1 and 2 show Western blotting analysis of the cell lysates probed with pan sodium channel antibody (top) and GFP (bottom) to show comparable levels of Na v 1.6/GFP (lane 1) and Na v 1.6/FHF2B-GFP (lane 2) in the samples used for the immunoprecipitation assay.

    Techniques Used: Negative Control, Binding Assay, Western Blot, Immunoprecipitation, Molecular Weight, Marker, Immunolabeling, Transfection

    13) Product Images from "Mbf1 ensures Polycomb silencing by protecting E(z) mRNA from degradation by Pacman"

    Article Title: Mbf1 ensures Polycomb silencing by protecting E(z) mRNA from degradation by Pacman

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.162461

    Conceivable functions of cytoplasmic Mbf1 protein via binding to mRNAs. (A) Model for Mbf1-ensured Polycomb silencing. In wild-type and mbf1 mutant lines, Pcm is not upregulated. Therefore, the steady-state level of E(z) mRNA is well balanced irrespective of Mbf1 expression. In Polycomb group mutants, Pcm expression is upregulated so that E(z) mRNA could become susceptible to Pcm attack. However, Mbf1 protects E(z) mRNA to ensure robustness of Polycomb silencing. In mbf1 and Polycomb group double mutants, loss of Mbf1 allows extensive degradation of E(z) mRNA by derepressed Pcm, thereby affecting Polycomb silencing. (B) The enrichment of four representative mRNAs ( GstD5 , Ide , Tep2 and Pebp1 ) identified in the RIP-seq results was confirmed by RIP RT-qPCR analysis. Results for E(z) and RpL30 mRNAs from Fig. 1 F are included for comparison. Data are mean±s.d. of fold-change versus control IgG; * P
    Figure Legend Snippet: Conceivable functions of cytoplasmic Mbf1 protein via binding to mRNAs. (A) Model for Mbf1-ensured Polycomb silencing. In wild-type and mbf1 mutant lines, Pcm is not upregulated. Therefore, the steady-state level of E(z) mRNA is well balanced irrespective of Mbf1 expression. In Polycomb group mutants, Pcm expression is upregulated so that E(z) mRNA could become susceptible to Pcm attack. However, Mbf1 protects E(z) mRNA to ensure robustness of Polycomb silencing. In mbf1 and Polycomb group double mutants, loss of Mbf1 allows extensive degradation of E(z) mRNA by derepressed Pcm, thereby affecting Polycomb silencing. (B) The enrichment of four representative mRNAs ( GstD5 , Ide , Tep2 and Pebp1 ) identified in the RIP-seq results was confirmed by RIP RT-qPCR analysis. Results for E(z) and RpL30 mRNAs from Fig. 1 F are included for comparison. Data are mean±s.d. of fold-change versus control IgG; * P

    Techniques Used: Binding Assay, Mutagenesis, Expressing, Quantitative RT-PCR

    14) Product Images from "Control of somatic tissue differentiation by the long non-coding RNA TINCR"

    Article Title: Control of somatic tissue differentiation by the long non-coding RNA TINCR

    Journal: Nature

    doi: 10.1038/nature11661

    TINCR interacts with differentiation mRNAs and STAU1 protein a , Enriched GO terms in TINCR-interacting genes detected by RIA-Seq. b , Protein microarray analysis detects TINCR RNA binding to STAU1 protein. Human recombinant protein microarray spotted with approximately 9,400 proteins (left); enlarged 144 protein spot subarray (middle) demonstrating strand-specific binding of TINCR sense strand to STAU1 protein (right); DUPD1 protein negative control is shown. Alexa-Fluor-647-labelled rabbit anti-mouse IgG in the top left corner of each subarray. c , STAU1 protein immunoprecipitation pulls down TINCR RNA. ANCR and LINC1 (also known as XIST ) represent lncRNA controls. d , Streptavidin precipitation of in vitro synthesized biotinylated TINCR RNA pulls down STAU1 protein. HA, haemagglutinin; WB, western blot.
    Figure Legend Snippet: TINCR interacts with differentiation mRNAs and STAU1 protein a , Enriched GO terms in TINCR-interacting genes detected by RIA-Seq. b , Protein microarray analysis detects TINCR RNA binding to STAU1 protein. Human recombinant protein microarray spotted with approximately 9,400 proteins (left); enlarged 144 protein spot subarray (middle) demonstrating strand-specific binding of TINCR sense strand to STAU1 protein (right); DUPD1 protein negative control is shown. Alexa-Fluor-647-labelled rabbit anti-mouse IgG in the top left corner of each subarray. c , STAU1 protein immunoprecipitation pulls down TINCR RNA. ANCR and LINC1 (also known as XIST ) represent lncRNA controls. d , Streptavidin precipitation of in vitro synthesized biotinylated TINCR RNA pulls down STAU1 protein. HA, haemagglutinin; WB, western blot.

    Techniques Used: Microarray, RNA Binding Assay, Recombinant, Binding Assay, Negative Control, Immunoprecipitation, In Vitro, Synthesized, Western Blot

    15) Product Images from "Expression of Zfhep/?EF1 protein in palate, neural progenitors, and differentiated neurons"

    Article Title: Expression of Zfhep/?EF1 protein in palate, neural progenitors, and differentiated neurons

    Journal: Gene expression patterns : GEP

    doi:

    Zfhep expression in the telencephalon. (A) Anti-Zfhep (1:400 dilution) and goat FITC-anti-rabbit IgG secondary antibody immunofluorescence of a transverse cryosection of the forebrain of an E14.5 embryo. The bar is 300 microns. (B) E13.5 mouse embryo brain labeled with anti-Zfhep showing the anterior portion of the superior horn of a lateral ventricle (LV) with Zfhep expression in cells of the ventricular zone (VZ). The bar is 40 microns. (C) Matched exposure of a section adjacent to B, and incubated with preimmune serum and FITC-secondary Ab, indicating the specificity of the anti-Zfhep antiserum. (D) Transverse cryostat section of lateral ventricle of E13.5 embryo double-labeled with monoclonal anti-neurofilament 165 (detected with FITC-secondary Ab) and anti-Zfhep (detected with donkey anti-rabbit IgG-rhodamine). The VZ contains Zfhep-positive (red) nuclei, however, cells that have differentiated are neurofilament-positive (green neurites) and Zfhep-negative. Bar is 40 microns. (E) LV of an E14.5 brain labeled with anti-Zfhep and rhodamine. Bar is 100 microns. (F) The same section labeled with anti-PCNA and FITC. (G) Combination of F and G showing co-expression of Zfhep and PCNA. *, 3 rd ventricle.
    Figure Legend Snippet: Zfhep expression in the telencephalon. (A) Anti-Zfhep (1:400 dilution) and goat FITC-anti-rabbit IgG secondary antibody immunofluorescence of a transverse cryosection of the forebrain of an E14.5 embryo. The bar is 300 microns. (B) E13.5 mouse embryo brain labeled with anti-Zfhep showing the anterior portion of the superior horn of a lateral ventricle (LV) with Zfhep expression in cells of the ventricular zone (VZ). The bar is 40 microns. (C) Matched exposure of a section adjacent to B, and incubated with preimmune serum and FITC-secondary Ab, indicating the specificity of the anti-Zfhep antiserum. (D) Transverse cryostat section of lateral ventricle of E13.5 embryo double-labeled with monoclonal anti-neurofilament 165 (detected with FITC-secondary Ab) and anti-Zfhep (detected with donkey anti-rabbit IgG-rhodamine). The VZ contains Zfhep-positive (red) nuclei, however, cells that have differentiated are neurofilament-positive (green neurites) and Zfhep-negative. Bar is 40 microns. (E) LV of an E14.5 brain labeled with anti-Zfhep and rhodamine. Bar is 100 microns. (F) The same section labeled with anti-PCNA and FITC. (G) Combination of F and G showing co-expression of Zfhep and PCNA. *, 3 rd ventricle.

    Techniques Used: Expressing, Immunofluorescence, Labeling, Incubation

    ), plate 32b-c) labeled with anti-Zfhep (detected with FITC) and anti-neurofilament 165 (detected with anti-mouse IgG-rhodamine). The labyrinth of the inner ear is to the left. Labeled neurofilament (red) shows differentiation of cells in the pons and in the vestibulocochlear (VIII) ganglion. Zfhep (green) strongly labels the anterior PEMS. 4 th , 4 th ventricle; CP, choroid plexus; MO, medulla oblongata; P, pons; PEMS, anterior precerebellar extramural migratory stream; PCN, precerebellar neuroepithelium; rV, proximal rootlets of the trigeminal nerve.
    Figure Legend Snippet: ), plate 32b-c) labeled with anti-Zfhep (detected with FITC) and anti-neurofilament 165 (detected with anti-mouse IgG-rhodamine). The labyrinth of the inner ear is to the left. Labeled neurofilament (red) shows differentiation of cells in the pons and in the vestibulocochlear (VIII) ganglion. Zfhep (green) strongly labels the anterior PEMS. 4 th , 4 th ventricle; CP, choroid plexus; MO, medulla oblongata; P, pons; PEMS, anterior precerebellar extramural migratory stream; PCN, precerebellar neuroepithelium; rV, proximal rootlets of the trigeminal nerve.

    Techniques Used: Labeling

    16) Product Images from "Generation and Characterization of a Diabody Targeting the ?v?6 Integrin"

    Article Title: Generation and Characterization of a Diabody Targeting the ?v?6 Integrin

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0073260

    Treatment of α v β 6 -expressing cells with B6.3 diabody resulted in diabody internalization and blockade of integrin functions. A) Localization of B6.3 diabody in A375Pβ6 cells by confocal microscopy. B6.3 diabody detection showed membrane pattern of staining at 4°C and internalized when cells were incubated at 37°C for 30 min, 1 h and 3 h. B6.3 diabody was detected using rabbit anti-human IgG followed by Alexa Fluor 546®-labeled goat anti-rabbit IgG (red). Cells were also counterstained with Hoechst 33245 (blue). B) Treatment of α v β 6 -expressing cells blocked adhesion to LAP-coated plates (A375Pβ6 and Capan-1 cells) and/or fibronectin-coated plates (Capan-1 cells). Cells were incubated with B6.3 or shMFE23 diabody at 4°C for 1 h and allowed to attach to coated plates for 1 h at 37°C. Treatment with the anti-CEA shMFE23 diabody had no effect on the cell lines used. C) B6.3 diabody treatment inhibited migration towards LAP and fibronectin. As observed in adhesion assays, the diabody inhibited migration of A375Pβ6 cells to LAP and migration of Capan-1 cells to fibronectin and LAP, while targeting CEA had no effect on the cells tested.
    Figure Legend Snippet: Treatment of α v β 6 -expressing cells with B6.3 diabody resulted in diabody internalization and blockade of integrin functions. A) Localization of B6.3 diabody in A375Pβ6 cells by confocal microscopy. B6.3 diabody detection showed membrane pattern of staining at 4°C and internalized when cells were incubated at 37°C for 30 min, 1 h and 3 h. B6.3 diabody was detected using rabbit anti-human IgG followed by Alexa Fluor 546®-labeled goat anti-rabbit IgG (red). Cells were also counterstained with Hoechst 33245 (blue). B) Treatment of α v β 6 -expressing cells blocked adhesion to LAP-coated plates (A375Pβ6 and Capan-1 cells) and/or fibronectin-coated plates (Capan-1 cells). Cells were incubated with B6.3 or shMFE23 diabody at 4°C for 1 h and allowed to attach to coated plates for 1 h at 37°C. Treatment with the anti-CEA shMFE23 diabody had no effect on the cell lines used. C) B6.3 diabody treatment inhibited migration towards LAP and fibronectin. As observed in adhesion assays, the diabody inhibited migration of A375Pβ6 cells to LAP and migration of Capan-1 cells to fibronectin and LAP, while targeting CEA had no effect on the cells tested.

    Techniques Used: Expressing, Confocal Microscopy, Staining, Incubation, Labeling, Migration

    B6.3 diabody inhibited LAP-mediated Smad2/3 translocation to the nucleus in Capan-1 cells. Cells were incubated at 4°C in the presence of B6.3 diabody and then treated with LAP or TGFβ 1 (30 min, 37°C); Smad2/3 localization was assessed by confocal microscopy (40X) using rabbit anti-Smad2/3 followed by Alexa Fluor 488®-labeled goat anti-rabbit IgG (green); smad2/3 was found in the cytoplasm of starved Capan-1 cells (a) and after treatment with B6.3 diabody (c). Smad2/3 was present in the nuclei in response to treatment with latent TGFβ 1 (b), a translocation that was inhibited by pre-treatment B6.3 diabody (d). TGFβ 1 was used as a positive control (e.f).
    Figure Legend Snippet: B6.3 diabody inhibited LAP-mediated Smad2/3 translocation to the nucleus in Capan-1 cells. Cells were incubated at 4°C in the presence of B6.3 diabody and then treated with LAP or TGFβ 1 (30 min, 37°C); Smad2/3 localization was assessed by confocal microscopy (40X) using rabbit anti-Smad2/3 followed by Alexa Fluor 488®-labeled goat anti-rabbit IgG (green); smad2/3 was found in the cytoplasm of starved Capan-1 cells (a) and after treatment with B6.3 diabody (c). Smad2/3 was present in the nuclei in response to treatment with latent TGFβ 1 (b), a translocation that was inhibited by pre-treatment B6.3 diabody (d). TGFβ 1 was used as a positive control (e.f).

    Techniques Used: Translocation Assay, Incubation, Confocal Microscopy, Labeling, Positive Control

    17) Product Images from "Epstein–Barr Virus Lytic Reactivation Induces IgG4 Production by Host B Lymphocytes in Graves' Disease Patients and Controls: A Subset of Graves' Disease Is an IgG4-Related Disease-Like Condition"

    Article Title: Epstein–Barr Virus Lytic Reactivation Induces IgG4 Production by Host B Lymphocytes in Graves' Disease Patients and Controls: A Subset of Graves' Disease Is an IgG4-Related Disease-Like Condition

    Journal: Viral Immunology

    doi: 10.1089/vim.2018.0042

    Detection of IgG4(+)72A1(+) double-positive cells in culture cells on day 5. We detected IgG4-positive and EBV-reactivated [IgG4(+)72A1(+)] cells in culture cells on day 5 and confirmed sorted cells by confocal laser microscope. Red spots are surface IgG4 and fine green dots are 72A1 antibodies binding to EBV-gp350/220. The frequency of 72A1(+) cells of this sample (representative case; patient 3) was 4.13% (0.67 + 3.46), and the mean frequency of four samples was 9.88% ( Supplementary Table S1 ).
    Figure Legend Snippet: Detection of IgG4(+)72A1(+) double-positive cells in culture cells on day 5. We detected IgG4-positive and EBV-reactivated [IgG4(+)72A1(+)] cells in culture cells on day 5 and confirmed sorted cells by confocal laser microscope. Red spots are surface IgG4 and fine green dots are 72A1 antibodies binding to EBV-gp350/220. The frequency of 72A1(+) cells of this sample (representative case; patient 3) was 4.13% (0.67 + 3.46), and the mean frequency of four samples was 9.88% ( Supplementary Table S1 ).

    Techniques Used: Microscopy, Binding Assay

    Production of IgG and IgG4 during the EBV reactivation period. Culture fluids were sampled on days 0, 5, 10, and 12, and IgG (A) and IgG4 (B) were then measured by ELISA. Time course changes were significant in Friedman's analysis of variance. IgG4 percentages (C) were higher than normal serum levels (approximately 4%). However, no significant difference was observed between patients and controls (D) .
    Figure Legend Snippet: Production of IgG and IgG4 during the EBV reactivation period. Culture fluids were sampled on days 0, 5, 10, and 12, and IgG (A) and IgG4 (B) were then measured by ELISA. Time course changes were significant in Friedman's analysis of variance. IgG4 percentages (C) were higher than normal serum levels (approximately 4%). However, no significant difference was observed between patients and controls (D) .

    Techniques Used: Enzyme-linked Immunosorbent Assay

    EBER1-positive cells and IgG4-positive cells were observed in the same areas. Resected tissues showed the diffuse hyperplasia of thyroid follicular epithelial cells with the focal infiltration of lymphocytes, but not tumefactive lesions, storiform fibrosis, or obliterative phlebitis (A) . EBER1-positive cells and IgG4-positive plasma cells were observed in the same area with lymphoid cell infiltration (B, D) . Six of the seven cases had a large number of IgG4-positive plasma cells (10/HPF
    Figure Legend Snippet: EBER1-positive cells and IgG4-positive cells were observed in the same areas. Resected tissues showed the diffuse hyperplasia of thyroid follicular epithelial cells with the focal infiltration of lymphocytes, but not tumefactive lesions, storiform fibrosis, or obliterative phlebitis (A) . EBER1-positive cells and IgG4-positive plasma cells were observed in the same area with lymphoid cell infiltration (B, D) . Six of the seven cases had a large number of IgG4-positive plasma cells (10/HPF

    Techniques Used:

    18) Product Images from "Tetraspanin CD82 interaction with cholesterol promotes extracellular vesicle–mediated release of ezrin to inhibit tumour cell movement"

    Article Title: Tetraspanin CD82 interaction with cholesterol promotes extracellular vesicle–mediated release of ezrin to inhibit tumour cell movement

    Journal: Journal of Extracellular Vesicles

    doi: 10.1080/20013078.2019.1692417

    CD82 and its cholesterol-binding differentially regulate cellular release of EVs. (a) Extracellular staining by filipin and Alexa488-conjugated CTxb in Du145 transfectants. Equal number of the cells were cultured on glass coverslips for 2 days, then fixed and labelled with filipin or Alexa488-conjugated CTxb. For filipin staining, intercellular regions were imaged. For CTxb staining, pericellular regions were imaged. Scale bar: 10 µm. (b) Distributions of Annexin V and Annexin A2 in Du145 transfectant cells. Alexa488-conjugated recombinant Annexin V was used for phosphatidylserine labelling, while Annexin-A2 Ab was used for Annexin-A2 staining. Scale bar: 10 μm. (c) Colocalization of Ezrin with GM1 or Annexin A2 in EVs. For Ezrin and GM1 co-staining, the cells were labelled with the Abs, Alexa488-conjugated CTxB and DAPI. For Ezrin and Annexin A2 co-staining, the cells were incubated sequentially with the primary Abs, Cy3-conjugated donkey anti-goat IgG, normal goat IgG and Alexa594-conjugated goat anti-mouse IgG. Images were obtained by confocal microscopy. Scale bar: 10 µm. (d) The cells were seeded in six-well plate at 50% confluence and cultured in DMEM containing 1% exosome-depleted FBS for 2 – 3 days. The culture supernatants were collected, spun at 2000 × g for 10 min to remove cell debris, and then analysed with NanoSight instrument for EV number and size. Data are presented as mean ± SD (n = 3 individual experiments). *: p
    Figure Legend Snippet: CD82 and its cholesterol-binding differentially regulate cellular release of EVs. (a) Extracellular staining by filipin and Alexa488-conjugated CTxb in Du145 transfectants. Equal number of the cells were cultured on glass coverslips for 2 days, then fixed and labelled with filipin or Alexa488-conjugated CTxb. For filipin staining, intercellular regions were imaged. For CTxb staining, pericellular regions were imaged. Scale bar: 10 µm. (b) Distributions of Annexin V and Annexin A2 in Du145 transfectant cells. Alexa488-conjugated recombinant Annexin V was used for phosphatidylserine labelling, while Annexin-A2 Ab was used for Annexin-A2 staining. Scale bar: 10 μm. (c) Colocalization of Ezrin with GM1 or Annexin A2 in EVs. For Ezrin and GM1 co-staining, the cells were labelled with the Abs, Alexa488-conjugated CTxB and DAPI. For Ezrin and Annexin A2 co-staining, the cells were incubated sequentially with the primary Abs, Cy3-conjugated donkey anti-goat IgG, normal goat IgG and Alexa594-conjugated goat anti-mouse IgG. Images were obtained by confocal microscopy. Scale bar: 10 µm. (d) The cells were seeded in six-well plate at 50% confluence and cultured in DMEM containing 1% exosome-depleted FBS for 2 – 3 days. The culture supernatants were collected, spun at 2000 × g for 10 min to remove cell debris, and then analysed with NanoSight instrument for EV number and size. Data are presented as mean ± SD (n = 3 individual experiments). *: p

    Techniques Used: Binding Assay, Staining, Cell Culture, Transfection, Recombinant, Incubation, Confocal Microscopy

    19) Product Images from "FXYD8, a Novel Regulator of Renal Na+/K+-ATPase in the Euryhaline Teleost, Tetraodon nigroviridis"

    Article Title: FXYD8, a Novel Regulator of Renal Na+/K+-ATPase in the Euryhaline Teleost, Tetraodon nigroviridis

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2017.00576

    Co-immunoprecipitation (Co-IP) of TnFXYD8 and NKA α-subunit (NKA α) in the pufferfish kidneys. TnFXYD8 or NKA were immunoprecipitated from renal crude membrane fractions of freshwater pufferfish using primary antibodies, then the immune complexes were analyzed by immunoblotting for NKA α (A) or TnFXYD8 (B) , respectively. In immunoprecipitated NKA α or TnFXYD8, the immunoblotting analyses for NKAα and TnFXYD8 revealed immunoreactive bands at 100 kDa (A) and 13 kDa (B) , respectively. In ( B) , the 55-kDa band in lane 3 is the IgG heavy chain of TnFXYD8 antibody. M, marker (kDa); lane 1, immunoblot detection of the opposing antibody (experimental group); lane 2, negative control for no antibody incubation in IP; lane 3, positive control using the same antibody with IP.
    Figure Legend Snippet: Co-immunoprecipitation (Co-IP) of TnFXYD8 and NKA α-subunit (NKA α) in the pufferfish kidneys. TnFXYD8 or NKA were immunoprecipitated from renal crude membrane fractions of freshwater pufferfish using primary antibodies, then the immune complexes were analyzed by immunoblotting for NKA α (A) or TnFXYD8 (B) , respectively. In immunoprecipitated NKA α or TnFXYD8, the immunoblotting analyses for NKAα and TnFXYD8 revealed immunoreactive bands at 100 kDa (A) and 13 kDa (B) , respectively. In ( B) , the 55-kDa band in lane 3 is the IgG heavy chain of TnFXYD8 antibody. M, marker (kDa); lane 1, immunoblot detection of the opposing antibody (experimental group); lane 2, negative control for no antibody incubation in IP; lane 3, positive control using the same antibody with IP.

    Techniques Used: Immunoprecipitation, Co-Immunoprecipitation Assay, Marker, Negative Control, Incubation, Positive Control

    20) Product Images from "Intronic Deletions That Disrupt mRNA Splicing of the tva Receptor Gene Result in Decreased Susceptibility to Infection by Avian Sarcoma and Leukosis Virus Subgroup A"

    Article Title: Intronic Deletions That Disrupt mRNA Splicing of the tva Receptor Gene Result in Decreased Susceptibility to Infection by Avian Sarcoma and Leukosis Virus Subgroup A

    Journal: Journal of Virology

    doi: 10.1128/JVI.05771-11

    Comparison of the Tva display on the surface of CEFs from CB, L15, and P chicken lines. Cells were incubated with crude extracellular supernatants containing the SU(A)-rIgG immunoadhesin and with goat-anti-rabbit IgG conjugated with Alexa Fluor 488. The
    Figure Legend Snippet: Comparison of the Tva display on the surface of CEFs from CB, L15, and P chicken lines. Cells were incubated with crude extracellular supernatants containing the SU(A)-rIgG immunoadhesin and with goat-anti-rabbit IgG conjugated with Alexa Fluor 488. The

    Techniques Used: Incubation

    21) Product Images from "The urokinase receptor-derived cyclic peptide [SRSRY] suppresses neovascularization and intravasation of osteosarcoma and chondrosarcoma cells"

    Article Title: The urokinase receptor-derived cyclic peptide [SRSRY] suppresses neovascularization and intravasation of osteosarcoma and chondrosarcoma cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.9976

    Inhibitory effect of [SRSRY] on migration of FPR1expressing osteosarcoma and chondrosarcoma cells ( A–B ) Representative images of human osteosarcoma Saos-2 and MG-63 cells, and human chondrosarcoma Sarc cells incubated with 2 μg/mL R4 anti-uPAR monoclonal antibody (A) or 1:100 anti-FPR1 polyclonal antibody 2 h at 23°C, exposed to Alexa 488-coniugated F(ab')2 fragment of rabbit anti-mouse IgG or goat anti-rabbit IgG for 40 min at 23°C and visualized by a fluorescence inverted microscope. Nuclei were stained blue with DAPI. Scale bar: 10 μm. Original magnification: 1000 x. ( C–D ) Whole cell lysates (20 and 40 μg/sample) from Saos-2, MG-63 and Sarc cells were resolved on a 10% SDS-PAGE under unreducing (C) or reducing conditions (D), followed by Western blotting with 1 μg/mL R4 anti-uPAR monoclonal antibody (C) or 1 μg/mL anti-FPR1 polyclonal antibody (D) and 0.2 μg/mL anti-GAPDH polyclonal antibody as loading control. The enclosed bar graphs show the average quantification of the uPAR/GAPDH (C) and FPR1/GAPDH (D) content from 3 independent experiments. ( E–F ) Saos-2, MG-63 and Sarc cells were allowed to migrate for 4 h at 37°C in 5% CO 2 in Boyden chambers toward DMEM (CTRL), or 10 nM [SRSRY] (E), DMEM (CTRL) or 10 nM SRSRY, in the absence (None) or the presence of 10 nM [SRSRY] (F). In all cases, the extent of cell migration was expressed as a percentage of the basal cell migration assessed toward serum-free medium, considered as 100% (CTRL). Data are expressed as the mean ± SD of three independent experiments, performed in triplicate. ***Statistical significance calculated against the positive control (None) with p
    Figure Legend Snippet: Inhibitory effect of [SRSRY] on migration of FPR1expressing osteosarcoma and chondrosarcoma cells ( A–B ) Representative images of human osteosarcoma Saos-2 and MG-63 cells, and human chondrosarcoma Sarc cells incubated with 2 μg/mL R4 anti-uPAR monoclonal antibody (A) or 1:100 anti-FPR1 polyclonal antibody 2 h at 23°C, exposed to Alexa 488-coniugated F(ab')2 fragment of rabbit anti-mouse IgG or goat anti-rabbit IgG for 40 min at 23°C and visualized by a fluorescence inverted microscope. Nuclei were stained blue with DAPI. Scale bar: 10 μm. Original magnification: 1000 x. ( C–D ) Whole cell lysates (20 and 40 μg/sample) from Saos-2, MG-63 and Sarc cells were resolved on a 10% SDS-PAGE under unreducing (C) or reducing conditions (D), followed by Western blotting with 1 μg/mL R4 anti-uPAR monoclonal antibody (C) or 1 μg/mL anti-FPR1 polyclonal antibody (D) and 0.2 μg/mL anti-GAPDH polyclonal antibody as loading control. The enclosed bar graphs show the average quantification of the uPAR/GAPDH (C) and FPR1/GAPDH (D) content from 3 independent experiments. ( E–F ) Saos-2, MG-63 and Sarc cells were allowed to migrate for 4 h at 37°C in 5% CO 2 in Boyden chambers toward DMEM (CTRL), or 10 nM [SRSRY] (E), DMEM (CTRL) or 10 nM SRSRY, in the absence (None) or the presence of 10 nM [SRSRY] (F). In all cases, the extent of cell migration was expressed as a percentage of the basal cell migration assessed toward serum-free medium, considered as 100% (CTRL). Data are expressed as the mean ± SD of three independent experiments, performed in triplicate. ***Statistical significance calculated against the positive control (None) with p

    Techniques Used: Migration, Incubation, Fluorescence, Inverted Microscopy, Staining, SDS Page, Western Blot, Positive Control

    22) Product Images from "Human Antibody Responses to a Chlamydia-Secreted Protease Factor "

    Article Title: Human Antibody Responses to a Chlamydia-Secreted Protease Factor

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.12.7164-7171.2004

    Detection of chlamydial protein-specific antibodies in human sera by Western blotting. The chlamydial GST fusion proteins, including GST-CPAF (lane 1), GST-MOMP (lane 2), and GST-HSP60 (lane 3), were resolved on an SDS-polyacrylamide gel and transferred onto nitrocellulose membranes for measuring human antibody binding to the corresponding chlamydial proteins. The human sera were pooled from 10 patients and subjected to fivefold serial dilution (as indicated at the top of the figure). The primary antibody binding was visualized with an HRP-conjugated goat anti-human IgG and ECL. The GST-CPAF band was obviously detected after 1:312,500 dilution of the human sera while the GST-MOMP and GST-HSP60 were no longer or minimally detected at the same serum dilution. The leftmost panel is an image of the gel stained with brilliant blue dye for total amounts of protein loaded to each lane. Lane 0 was loaded with a prestained molecular mass marker.
    Figure Legend Snippet: Detection of chlamydial protein-specific antibodies in human sera by Western blotting. The chlamydial GST fusion proteins, including GST-CPAF (lane 1), GST-MOMP (lane 2), and GST-HSP60 (lane 3), were resolved on an SDS-polyacrylamide gel and transferred onto nitrocellulose membranes for measuring human antibody binding to the corresponding chlamydial proteins. The human sera were pooled from 10 patients and subjected to fivefold serial dilution (as indicated at the top of the figure). The primary antibody binding was visualized with an HRP-conjugated goat anti-human IgG and ECL. The GST-CPAF band was obviously detected after 1:312,500 dilution of the human sera while the GST-MOMP and GST-HSP60 were no longer or minimally detected at the same serum dilution. The leftmost panel is an image of the gel stained with brilliant blue dye for total amounts of protein loaded to each lane. Lane 0 was loaded with a prestained molecular mass marker.

    Techniques Used: Western Blot, Binding Assay, Serial Dilution, Staining, Marker

    23) Product Images from "Regulatory Phosphorylation of Ikaros by Bruton's Tyrosine Kinase"

    Article Title: Regulatory Phosphorylation of Ikaros by Bruton's Tyrosine Kinase

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071302

    Co-localization and Physical Interactions of Native Ikaros and BTK Proteins in Human Cells. [A] Nuclear co-localization of Native IK and BTK. ALL-N1 cells were fixed and stained with polyclonal rabbit anti-IK1 (primary Ab)/Alexa Fluor 568 F(ab') 2 fragment of goat anti-rabbit IgG (secondary Ab) (red) and mouse anti-BTK MoAb (primary Ab)/Alexa Fluor 488 goat anti-mouse IgG (secondary Ab) (green) antibodies. Nuclei were stained with blue fluorescent dye 4′,6-diamidino-2-phenylindole (DAPI). MERGE panels depict the merge three-color confocal image showing co-localization of IK1 and BTK in DAPI-stained nucleus as magenta immunofluorescent foci (System magnification: 315×). Representative foci of colocalization are indicated with white arrowheads. [B] Co-immunoprecipitation of Native IK and BTK. B1 depicts the results of the BTK Western blot analysis of the IK and BTK immune complexes immunoprecipitated (IP) from ALL-N1 cells. B.2 depicts the results of the IK Western blot analysis of the BTK and IK immune complexes from the same cells. Controls included immunoprecipitations performed without using a primary (1 0 ) antibody.
    Figure Legend Snippet: Co-localization and Physical Interactions of Native Ikaros and BTK Proteins in Human Cells. [A] Nuclear co-localization of Native IK and BTK. ALL-N1 cells were fixed and stained with polyclonal rabbit anti-IK1 (primary Ab)/Alexa Fluor 568 F(ab') 2 fragment of goat anti-rabbit IgG (secondary Ab) (red) and mouse anti-BTK MoAb (primary Ab)/Alexa Fluor 488 goat anti-mouse IgG (secondary Ab) (green) antibodies. Nuclei were stained with blue fluorescent dye 4′,6-diamidino-2-phenylindole (DAPI). MERGE panels depict the merge three-color confocal image showing co-localization of IK1 and BTK in DAPI-stained nucleus as magenta immunofluorescent foci (System magnification: 315×). Representative foci of colocalization are indicated with white arrowheads. [B] Co-immunoprecipitation of Native IK and BTK. B1 depicts the results of the BTK Western blot analysis of the IK and BTK immune complexes immunoprecipitated (IP) from ALL-N1 cells. B.2 depicts the results of the IK Western blot analysis of the BTK and IK immune complexes from the same cells. Controls included immunoprecipitations performed without using a primary (1 0 ) antibody.

    Techniques Used: Staining, Immunoprecipitation, Western Blot

    BTK Expression Levels Control Nuclear Localization of Ikaros. [ A ] Cells were stained with mouse monoclonal antibody (mAb) against human BTK and an anti-IK mouse mAb, which was raised against murine IK and which cross-reacts with human IK. TOTO-3 was used for nuclear staining. Depicted are the confocal two-color (green/blue: BTK/TOTO-3 and IK/TOTO-3) merge images of abundantly BTK + (high BTK) leukemic cells from a pediatric B-precursor ALL case vs. leukemic cells with low BTK expression level from an infant B-precursor ALL case. Nuclear staining for IK was observed in the confocal fluorescence images of primary leukemic B-cell precursors with high BTK expression level. By contrast, leukemic cells with low BTK expression levels showed an aberrant, predominantly cytoplasmic localization of IK (System Magnification: 500×). [ B ] The DT-40 cell line and its subclones were stained with the rabbit polyclonal, H-100 (sc-13039) antibody against the N-terminus of IK. Depicted are the confocal two-color (red/blue) IK/DAPI merge images of wildtype (WT) DT40 cells, BTK-deficient DT40 cells, and BTK-deficient DT40 cells reconstituted with wildtype btk . The contour of the DAPI-stained nuclei (blue) was marked with a dotted line and shows a significant amount of IK protein (red) outside the nucleus of the BTK − DT40 cells. [ C1 ] Upper panel: Depicted are the confocal single-color and two-color merge images of control Hek293T cells stained with the secondary goat anti-mouse (GAM) antibody and DAPI. No false positive green fluorescence was detected (System Magnification: 630×). Lower panel: Depicted are the confocal single-color and two-color (green/blue: BTK/DAPI) merge images of test Hek293T cells stained with anti-BTK/goat-anti-mouse (GAM) antibody combination and DAPI. BTK (green fluorescence) was localized primarily in the cytoplasm of Hek293T cells around the DAPI-stained blue nucleus. [ C2 ] Left panel: Merge confocal images of untreated control Hek293T cells two-color stained with anti-BTK/goat-anti-mouse (GAM) antibody combination and DAPI. Right panel: Merge confocal images of BTK-siRNA transfected Hek293T cells two-color stained with anti-BTK/goat-anti-mouse (GAM) antibody combination and DAPI 72 h post-transfection. (System Magnification: 630×). BTK depletion was confirmed by Western blot analysis (see Figure 3 ) [ D ] Confocal images of Hek293T cells expressing the HA-tagged mutant IK proteins D 6 or A 12 stained with the mouse monoclonal anti-HA antibody (HA-probe F-7)(primary Ab)/Alexa Fluor 488 goat anti-mouse IgG (secondary Ab) (green) antibody and blue fluorescent DNA dye 4′, 6-diamidino-2-phenylindole (DAPI) following treatment with BTK siRNA (50 nM) vs. PBS (CON). (System Magnification: 630×).
    Figure Legend Snippet: BTK Expression Levels Control Nuclear Localization of Ikaros. [ A ] Cells were stained with mouse monoclonal antibody (mAb) against human BTK and an anti-IK mouse mAb, which was raised against murine IK and which cross-reacts with human IK. TOTO-3 was used for nuclear staining. Depicted are the confocal two-color (green/blue: BTK/TOTO-3 and IK/TOTO-3) merge images of abundantly BTK + (high BTK) leukemic cells from a pediatric B-precursor ALL case vs. leukemic cells with low BTK expression level from an infant B-precursor ALL case. Nuclear staining for IK was observed in the confocal fluorescence images of primary leukemic B-cell precursors with high BTK expression level. By contrast, leukemic cells with low BTK expression levels showed an aberrant, predominantly cytoplasmic localization of IK (System Magnification: 500×). [ B ] The DT-40 cell line and its subclones were stained with the rabbit polyclonal, H-100 (sc-13039) antibody against the N-terminus of IK. Depicted are the confocal two-color (red/blue) IK/DAPI merge images of wildtype (WT) DT40 cells, BTK-deficient DT40 cells, and BTK-deficient DT40 cells reconstituted with wildtype btk . The contour of the DAPI-stained nuclei (blue) was marked with a dotted line and shows a significant amount of IK protein (red) outside the nucleus of the BTK − DT40 cells. [ C1 ] Upper panel: Depicted are the confocal single-color and two-color merge images of control Hek293T cells stained with the secondary goat anti-mouse (GAM) antibody and DAPI. No false positive green fluorescence was detected (System Magnification: 630×). Lower panel: Depicted are the confocal single-color and two-color (green/blue: BTK/DAPI) merge images of test Hek293T cells stained with anti-BTK/goat-anti-mouse (GAM) antibody combination and DAPI. BTK (green fluorescence) was localized primarily in the cytoplasm of Hek293T cells around the DAPI-stained blue nucleus. [ C2 ] Left panel: Merge confocal images of untreated control Hek293T cells two-color stained with anti-BTK/goat-anti-mouse (GAM) antibody combination and DAPI. Right panel: Merge confocal images of BTK-siRNA transfected Hek293T cells two-color stained with anti-BTK/goat-anti-mouse (GAM) antibody combination and DAPI 72 h post-transfection. (System Magnification: 630×). BTK depletion was confirmed by Western blot analysis (see Figure 3 ) [ D ] Confocal images of Hek293T cells expressing the HA-tagged mutant IK proteins D 6 or A 12 stained with the mouse monoclonal anti-HA antibody (HA-probe F-7)(primary Ab)/Alexa Fluor 488 goat anti-mouse IgG (secondary Ab) (green) antibody and blue fluorescent DNA dye 4′, 6-diamidino-2-phenylindole (DAPI) following treatment with BTK siRNA (50 nM) vs. PBS (CON). (System Magnification: 630×).

    Techniques Used: Expressing, Staining, Fluorescence, Transfection, Western Blot, Mutagenesis

    24) Product Images from "The BAF (BRG1/BRM-Associated Factor) Chromatin-Remodeling Complex Exhibits Ethanol Sensitivity in Fetal Neural Progenitor Cells and Regulates Transcription at the Mir-9-2 Encoding Gene Locus"

    Article Title: The BAF (BRG1/BRM-Associated Factor) Chromatin-Remodeling Complex Exhibits Ethanol Sensitivity in Fetal Neural Progenitor Cells and Regulates Transcription at the Mir-9-2 Encoding Gene Locus

    Journal: Alcohol (Fayetteville, N.Y.)

    doi: 10.1016/j.alcohol.2017.01.003

    BRG1 protein expression in NSCs (a) Western immunoblot shows a single BRG1 band at ~238KD detected in the nuclear extract of neurosphere cultures but not in the cytoplasmic extract. (b) BRG1 protein levels in neurospheres after 0mg/dl, 120mg/dl and 320mg/dl ethanol treatment. Lower panel shows the corresponding immunoblot when probed for β-Actin as a loading control. (c) Bar graph, depicting the quantification of the western blot, shows that ethanol did not alter BRG1 protein expression, consistent with the lack of effect on BRG1 mRNA expression. The vertical axis shows the ratio of the band density of BRG1 to the ratio of the band density of β-Actin. Error bars indicates standard error of the mean. (d) Western blot analysis of BRG1 immunoprecipitation in cytoplasmic and nuclear cellular fractions probed with anti-BRG1 antibody. BRG1 is specifically precipitated with anti-BRG1 antibody (blue text), but not with an isotype-specific IgG control antibody (red text). Efficiency of anti-BRG1 immunoprecipitation is indicated by the relative depletion of BRG1 from the nuclear supernatant and enrichment in the immuno-precipitate (IP).
    Figure Legend Snippet: BRG1 protein expression in NSCs (a) Western immunoblot shows a single BRG1 band at ~238KD detected in the nuclear extract of neurosphere cultures but not in the cytoplasmic extract. (b) BRG1 protein levels in neurospheres after 0mg/dl, 120mg/dl and 320mg/dl ethanol treatment. Lower panel shows the corresponding immunoblot when probed for β-Actin as a loading control. (c) Bar graph, depicting the quantification of the western blot, shows that ethanol did not alter BRG1 protein expression, consistent with the lack of effect on BRG1 mRNA expression. The vertical axis shows the ratio of the band density of BRG1 to the ratio of the band density of β-Actin. Error bars indicates standard error of the mean. (d) Western blot analysis of BRG1 immunoprecipitation in cytoplasmic and nuclear cellular fractions probed with anti-BRG1 antibody. BRG1 is specifically precipitated with anti-BRG1 antibody (blue text), but not with an isotype-specific IgG control antibody (red text). Efficiency of anti-BRG1 immunoprecipitation is indicated by the relative depletion of BRG1 from the nuclear supernatant and enrichment in the immuno-precipitate (IP).

    Techniques Used: Expressing, Western Blot, Immunoprecipitation

    BRG1-containing complexes associate with both DNAse I-hypersensitive and insensitive sites on the miR-9-2 gene locus (a) Positional representation of POU5F1/Oct4, c-myc and REST transcription regulatory factor binding sites along the human pre-miR-9-2 gene locus identified with the UCSC genome browser ENCODE analysis hub track. Colored circles indicate locations for primer pairs for pri-miR-9-2 regions 1,2,3,4 and 5 and the pre-miR-9-2 coding region. (b) qPCR results of BRG1-ChIP from neurospheres. Primers were used to amplify 6 distinct positions along the pri-miR-9-2 gene locus and a genetically sparse region on mouse chromosome 6. Vertical axis shows fold change relative to an IgG pulldown control. Error bars indicate standard error of the mean. (c) qPCR of neurosphere DNA following digestion with 20, 60 and 120U of DNAse I at the 6 positions along the pri-miR-9-2 gene locus. The vertical axis shows fold amplification relative to DNAse I-untreated neurosphere DNA.
    Figure Legend Snippet: BRG1-containing complexes associate with both DNAse I-hypersensitive and insensitive sites on the miR-9-2 gene locus (a) Positional representation of POU5F1/Oct4, c-myc and REST transcription regulatory factor binding sites along the human pre-miR-9-2 gene locus identified with the UCSC genome browser ENCODE analysis hub track. Colored circles indicate locations for primer pairs for pri-miR-9-2 regions 1,2,3,4 and 5 and the pre-miR-9-2 coding region. (b) qPCR results of BRG1-ChIP from neurospheres. Primers were used to amplify 6 distinct positions along the pri-miR-9-2 gene locus and a genetically sparse region on mouse chromosome 6. Vertical axis shows fold change relative to an IgG pulldown control. Error bars indicate standard error of the mean. (c) qPCR of neurosphere DNA following digestion with 20, 60 and 120U of DNAse I at the 6 positions along the pri-miR-9-2 gene locus. The vertical axis shows fold amplification relative to DNAse I-untreated neurosphere DNA.

    Techniques Used: Binding Assay, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Amplification

    25) Product Images from "Cytoskeletal Association Is Important for Differential Targeting of Glucose Transporter Isoforms in Leishmania"

    Article Title: Cytoskeletal Association Is Important for Differential Targeting of Glucose Transporter Isoforms in Leishmania

    Journal: The Journal of Cell Biology

    doi:

    Double-label confocal laser scanning micrographs of Triton X-100–extracted L. enriettii promastigotes stained with anti-P1C and anti– α-tubulin. Cytoskeletons were fixed with methanol, stained with a 1:100 dilution of the anti-P1C antibody and a 1:500 dilution of the anti–α-tubulin antibody, and then with an FITC-conjugated anti–rabbit IgG ( P1C ) and a rhodamine-conjugated anti–mouse IgG ( α-tubulin ) secondary antibody. Cytoskeletons were examined by confocal microscopy using illumination at 488 nm to visualize the Pro-1 glucose transporter–complexed FITC antibody ( P1C ) or at 546 nm to visualize α-tubulin complexed with the rhodamine antibody ( α-tubulin ). Each micrograph represents a single 0.5-μm section through each field.
    Figure Legend Snippet: Double-label confocal laser scanning micrographs of Triton X-100–extracted L. enriettii promastigotes stained with anti-P1C and anti– α-tubulin. Cytoskeletons were fixed with methanol, stained with a 1:100 dilution of the anti-P1C antibody and a 1:500 dilution of the anti–α-tubulin antibody, and then with an FITC-conjugated anti–rabbit IgG ( P1C ) and a rhodamine-conjugated anti–mouse IgG ( α-tubulin ) secondary antibody. Cytoskeletons were examined by confocal microscopy using illumination at 488 nm to visualize the Pro-1 glucose transporter–complexed FITC antibody ( P1C ) or at 546 nm to visualize α-tubulin complexed with the rhodamine antibody ( α-tubulin ). Each micrograph represents a single 0.5-μm section through each field.

    Techniques Used: Staining, Confocal Microscopy

    Double-label confocal laser scanning micrographs of Triton X-100–extracted L. enriettii promastigotes transfected with plasmid encoding epitope-tagged iso-1 ( top ) or epitope-tagged iso-2 ( bottom ) and stained with the rabbit anti-GLUT2 antibody directed against the epitope tag ( GLUT2 ) and the murine anti–α-tubulin antibody ( α-tubulin ). Cytoskeletons were fixed with methanol, stained with 1:500 dilutions of the anti-GLUT2 antibody and anti-α-tubulin antibodies, and then with an FITC-conjugated anti–rabbit IgG ( GLUT2 ) and a rhodamine-conjugated anti–mouse IgG ( α-tubulin ) secondary antibody. Cytoskeletons were examined by confocal microscopy. Each micrograph represents a single 0.5-μm section through each field.
    Figure Legend Snippet: Double-label confocal laser scanning micrographs of Triton X-100–extracted L. enriettii promastigotes transfected with plasmid encoding epitope-tagged iso-1 ( top ) or epitope-tagged iso-2 ( bottom ) and stained with the rabbit anti-GLUT2 antibody directed against the epitope tag ( GLUT2 ) and the murine anti–α-tubulin antibody ( α-tubulin ). Cytoskeletons were fixed with methanol, stained with 1:500 dilutions of the anti-GLUT2 antibody and anti-α-tubulin antibodies, and then with an FITC-conjugated anti–rabbit IgG ( GLUT2 ) and a rhodamine-conjugated anti–mouse IgG ( α-tubulin ) secondary antibody. Cytoskeletons were examined by confocal microscopy. Each micrograph represents a single 0.5-μm section through each field.

    Techniques Used: Transfection, Plasmid Preparation, Staining, Confocal Microscopy

    26) Product Images from "A premature stop codon within the tvb receptor gene results in decreased susceptibility to infection by avian leukosis virus subgroups B, D, and E"

    Article Title: A premature stop codon within the tvb receptor gene results in decreased susceptibility to infection by avian leukosis virus subgroups B, D, and E

    Journal: Oncotarget

    doi: 10.18632/oncotarget.22512

    Binding affinities of ALV envelope glycoproteins for Tvb receptors expressed on the surfaces of CEFs of defined origin The tvb s1/s1 , tvb s1/r3 and tvb r3/r3 CEFs were incubated with the same amount of soluble forms of the SU-rIgG, SU(B)-rIgG (A) , SUD-rIgG (B) or SUE-rIgG (C) . The viral envelope glycoprotein-receptor complexes were bound to goat-anti-rabbit IgG conjugated with Alexa Fluor 488, and the amount of Alexa Fluor 488 bound to the cells was quantitated by FACS. Binding affinities of the envelope-receptor is given as the percentage of fluorescence-positive cells. The values shown are means ± standard deviation from three independent experiments. To determine the statistical significance of differences between binding of wt and the mutant Tvb receptors for all three soluble SU glycoproteins data sets, the Student's t tests were performed ( ** , P
    Figure Legend Snippet: Binding affinities of ALV envelope glycoproteins for Tvb receptors expressed on the surfaces of CEFs of defined origin The tvb s1/s1 , tvb s1/r3 and tvb r3/r3 CEFs were incubated with the same amount of soluble forms of the SU-rIgG, SU(B)-rIgG (A) , SUD-rIgG (B) or SUE-rIgG (C) . The viral envelope glycoprotein-receptor complexes were bound to goat-anti-rabbit IgG conjugated with Alexa Fluor 488, and the amount of Alexa Fluor 488 bound to the cells was quantitated by FACS. Binding affinities of the envelope-receptor is given as the percentage of fluorescence-positive cells. The values shown are means ± standard deviation from three independent experiments. To determine the statistical significance of differences between binding of wt and the mutant Tvb receptors for all three soluble SU glycoproteins data sets, the Student's t tests were performed ( ** , P

    Techniques Used: Binding Assay, Incubation, FACS, Fluorescence, Standard Deviation, Mutagenesis

    27) Product Images from "Intestinal phenotype in mice overexpressing a heparin-binding EGF-like growth factor transgene in enterocytes"

    Article Title: Intestinal phenotype in mice overexpressing a heparin-binding EGF-like growth factor transgene in enterocytes

    Journal: Growth factors (Chur, Switzerland)

    doi: 10.3109/08977190903407365

    Intestinal histology and morphometric analysis of HB-EGF TG and WT mice. (A) Intestinal histology. Shown are representative H E-stained sections of duodenum from WT, high expression TG mice, and low expression TG mice at 1 month and 5 months of age. The scale bar represents 100 μm. (B) Intestinal morphometric analysis. Villous length, villous width, crypt depth, and muscle thickness in H E-stained sections of duodenum, jejunum, and ileum from WT, high expression TG mice, and low expression TG mice were quantified using ImageJ 1.39U software. (C) Enterocyte immunostaining. Shown are jejunal sections from WT, high expression TG, and low expression TG mice immunostained with rabbit anti-E-cadherin and secondary goat anti-rabbit IgG conjugated with Alexa 560 (red). The nulei were conterstained with DAPI (blue). The scale bar represents 50 μm. (D) Enterocyte cell density. Columnar enterocyte cell densities were quantified by counting the number of cells in the distal 200 μm from the distal tips of the villi. Each graph bar represents quantification of 15 villous tips. All values shown represent mean ± SD. Statistical significance was determined by one-way ANOVA (repeated measures). * p
    Figure Legend Snippet: Intestinal histology and morphometric analysis of HB-EGF TG and WT mice. (A) Intestinal histology. Shown are representative H E-stained sections of duodenum from WT, high expression TG mice, and low expression TG mice at 1 month and 5 months of age. The scale bar represents 100 μm. (B) Intestinal morphometric analysis. Villous length, villous width, crypt depth, and muscle thickness in H E-stained sections of duodenum, jejunum, and ileum from WT, high expression TG mice, and low expression TG mice were quantified using ImageJ 1.39U software. (C) Enterocyte immunostaining. Shown are jejunal sections from WT, high expression TG, and low expression TG mice immunostained with rabbit anti-E-cadherin and secondary goat anti-rabbit IgG conjugated with Alexa 560 (red). The nulei were conterstained with DAPI (blue). The scale bar represents 50 μm. (D) Enterocyte cell density. Columnar enterocyte cell densities were quantified by counting the number of cells in the distal 200 μm from the distal tips of the villi. Each graph bar represents quantification of 15 villous tips. All values shown represent mean ± SD. Statistical significance was determined by one-way ANOVA (repeated measures). * p

    Techniques Used: Mouse Assay, Staining, Expressing, Software, Immunostaining

    28) Product Images from "Antioxidant defense by thioredoxin can occur independently of canonical thiol-disulfide oxidoreductase enzymatic activity"

    Article Title: Antioxidant defense by thioredoxin can occur independently of canonical thiol-disulfide oxidoreductase enzymatic activity

    Journal: Cell reports

    doi: 10.1016/j.celrep.2016.02.066

    SsrB is a substrate of thioredoxin-1 thiol-disulfide oxidoreductase-dependent and –independent activities Western blotting of SsrB-3XFLAG after lysates of Salmonella containing pTRXATAP plasmids were purified sequentially with IgG (1 st elution) and calmodulin (2 nd elution) (A). Detection of the purified TrxA-6His proteins bound to the full-length of GST-SsrB by anti-6His immunoblot analysis after pull-down. The GST protein was used as control (B). Molecular markers are indicated. A recombinant C-terminal domain of SsrB separated by PAGE gel electrophoresis was visualized by Coomassie brilliant blue staining (C). Where indicated, 25 µM of the C-terminal domain of SsrB were oxidized with 250 µM H 2 O 2 . Some of the specimens exposed to H 2 O 2 were treated with 25 µM of recombinant TrxA or TrxA C32A C35A. The sizes of SsrB monomers and dimers are indicated on the right. TrxA-6His and TrxA C32A C35A-6His proteins were visualized by Western blot using an anti-6His antibody after the pull-down with recombinant GST-SsrB (D). Interactions between wild-type or TrxA C32A C35A with full-length SsrB were studied in a bacterial two-hybrid system that reconstitutes the T18 and T25 domains of adenylate cyclase (E). Thioredoxin reductase (TrxB) and the RpoA α-subunit of the RNA polymerase were included as positive and negative controls, respectively. The activity of reconstituted adenylate cyclase is expressed in Miller units (M.U.). Abundance of TrxA in soluble and whole cell cytoplasmic extracts of stationary phase Salmonella was determined by Western blotting (F). The amount of ssrB mRNA was quantified in overnight cultures of WT and Δ trxA Salmonella (G). The data are expressed relative to the rpoD .
    Figure Legend Snippet: SsrB is a substrate of thioredoxin-1 thiol-disulfide oxidoreductase-dependent and –independent activities Western blotting of SsrB-3XFLAG after lysates of Salmonella containing pTRXATAP plasmids were purified sequentially with IgG (1 st elution) and calmodulin (2 nd elution) (A). Detection of the purified TrxA-6His proteins bound to the full-length of GST-SsrB by anti-6His immunoblot analysis after pull-down. The GST protein was used as control (B). Molecular markers are indicated. A recombinant C-terminal domain of SsrB separated by PAGE gel electrophoresis was visualized by Coomassie brilliant blue staining (C). Where indicated, 25 µM of the C-terminal domain of SsrB were oxidized with 250 µM H 2 O 2 . Some of the specimens exposed to H 2 O 2 were treated with 25 µM of recombinant TrxA or TrxA C32A C35A. The sizes of SsrB monomers and dimers are indicated on the right. TrxA-6His and TrxA C32A C35A-6His proteins were visualized by Western blot using an anti-6His antibody after the pull-down with recombinant GST-SsrB (D). Interactions between wild-type or TrxA C32A C35A with full-length SsrB were studied in a bacterial two-hybrid system that reconstitutes the T18 and T25 domains of adenylate cyclase (E). Thioredoxin reductase (TrxB) and the RpoA α-subunit of the RNA polymerase were included as positive and negative controls, respectively. The activity of reconstituted adenylate cyclase is expressed in Miller units (M.U.). Abundance of TrxA in soluble and whole cell cytoplasmic extracts of stationary phase Salmonella was determined by Western blotting (F). The amount of ssrB mRNA was quantified in overnight cultures of WT and Δ trxA Salmonella (G). The data are expressed relative to the rpoD .

    Techniques Used: Western Blot, Purification, Recombinant, Polyacrylamide Gel Electrophoresis, Nucleic Acid Electrophoresis, Staining, Activity Assay

    29) Product Images from "Generation and Characterization of a Diabody Targeting the ?v?6 Integrin"

    Article Title: Generation and Characterization of a Diabody Targeting the ?v?6 Integrin

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0073260

    Treatment of α v β 6 -expressing cells with B6.3 diabody resulted in diabody internalization and blockade of integrin functions. A) Localization of B6.3 diabody in A375Pβ6 cells by confocal microscopy. B6.3 diabody detection showed membrane pattern of staining at 4°C and internalized when cells were incubated at 37°C for 30 min, 1 h and 3 h. B6.3 diabody was detected using rabbit anti-human IgG followed by Alexa Fluor 546®-labeled goat anti-rabbit IgG (red). Cells were also counterstained with Hoechst 33245 (blue). B) Treatment of α v β 6 -expressing cells blocked adhesion to LAP-coated plates (A375Pβ6 and Capan-1 cells) and/or fibronectin-coated plates (Capan-1 cells). Cells were incubated with B6.3 or shMFE23 diabody at 4°C for 1 h and allowed to attach to coated plates for 1 h at 37°C. Treatment with the anti-CEA shMFE23 diabody had no effect on the cell lines used. C) B6.3 diabody treatment inhibited migration towards LAP and fibronectin. As observed in adhesion assays, the diabody inhibited migration of A375Pβ6 cells to LAP and migration of Capan-1 cells to fibronectin and LAP, while targeting CEA had no effect on the cells tested.
    Figure Legend Snippet: Treatment of α v β 6 -expressing cells with B6.3 diabody resulted in diabody internalization and blockade of integrin functions. A) Localization of B6.3 diabody in A375Pβ6 cells by confocal microscopy. B6.3 diabody detection showed membrane pattern of staining at 4°C and internalized when cells were incubated at 37°C for 30 min, 1 h and 3 h. B6.3 diabody was detected using rabbit anti-human IgG followed by Alexa Fluor 546®-labeled goat anti-rabbit IgG (red). Cells were also counterstained with Hoechst 33245 (blue). B) Treatment of α v β 6 -expressing cells blocked adhesion to LAP-coated plates (A375Pβ6 and Capan-1 cells) and/or fibronectin-coated plates (Capan-1 cells). Cells were incubated with B6.3 or shMFE23 diabody at 4°C for 1 h and allowed to attach to coated plates for 1 h at 37°C. Treatment with the anti-CEA shMFE23 diabody had no effect on the cell lines used. C) B6.3 diabody treatment inhibited migration towards LAP and fibronectin. As observed in adhesion assays, the diabody inhibited migration of A375Pβ6 cells to LAP and migration of Capan-1 cells to fibronectin and LAP, while targeting CEA had no effect on the cells tested.

    Techniques Used: Expressing, Confocal Microscopy, Staining, Incubation, Labeling, Migration

    B6.3 diabody inhibited LAP-mediated Smad2/3 translocation to the nucleus in Capan-1 cells. Cells were incubated at 4°C in the presence of B6.3 diabody and then treated with LAP or TGFβ 1 (30 min, 37°C); Smad2/3 localization was assessed by confocal microscopy (40X) using rabbit anti-Smad2/3 followed by Alexa Fluor 488®-labeled goat anti-rabbit IgG (green); smad2/3 was found in the cytoplasm of starved Capan-1 cells (a) and after treatment with B6.3 diabody (c). Smad2/3 was present in the nuclei in response to treatment with latent TGFβ 1 (b), a translocation that was inhibited by pre-treatment B6.3 diabody (d). TGFβ 1 was used as a positive control (e.f).
    Figure Legend Snippet: B6.3 diabody inhibited LAP-mediated Smad2/3 translocation to the nucleus in Capan-1 cells. Cells were incubated at 4°C in the presence of B6.3 diabody and then treated with LAP or TGFβ 1 (30 min, 37°C); Smad2/3 localization was assessed by confocal microscopy (40X) using rabbit anti-Smad2/3 followed by Alexa Fluor 488®-labeled goat anti-rabbit IgG (green); smad2/3 was found in the cytoplasm of starved Capan-1 cells (a) and after treatment with B6.3 diabody (c). Smad2/3 was present in the nuclei in response to treatment with latent TGFβ 1 (b), a translocation that was inhibited by pre-treatment B6.3 diabody (d). TGFβ 1 was used as a positive control (e.f).

    Techniques Used: Translocation Assay, Incubation, Confocal Microscopy, Labeling, Positive Control

    30) Product Images from "Morphological and Functional Analyses of the Tight Junction in the Palatal Epithelium of Mouse"

    Article Title: Morphological and Functional Analyses of the Tight Junction in the Palatal Epithelium of Mouse

    Journal: Acta Histochemica et Cytochemica

    doi: 10.1267/ahc.17006

    Immunohistochemistry of TJ constitutive proteins. The immunofluorescence of OCD ( A ), CLD-1 ( C ), and -4 ( D ) were localized between the upper layers of the stratum granulosum showing dot-like fluorescence (arrowheads). In addition, CLD-1 ( C ) and CLD-4 ( D ) showed network-like positive reaction from the lower layer of the stratum granulosum to the upper layer of the stratum spinosum. CLD-2 ( F ) and CLD-5 ( G ) were positive in the salivary glands and the endothelium of blood vessel respectively, but were negative in the palatal epithelium. Background staining levels were checked by omitting the primary antibodies ( B : rabbit anti-goat IgG labeled with Alexa Fluor 488, E : goat anti-rabbit IgG labeled with Alexa Fluor 488). SC: stratum corneum, SB: stratum basale, Blue: nuclei stained with Hoechst. Bars = 25 μm ( A, B, C, D, E ), 50 μm ( F ), 10 μm ( G ).
    Figure Legend Snippet: Immunohistochemistry of TJ constitutive proteins. The immunofluorescence of OCD ( A ), CLD-1 ( C ), and -4 ( D ) were localized between the upper layers of the stratum granulosum showing dot-like fluorescence (arrowheads). In addition, CLD-1 ( C ) and CLD-4 ( D ) showed network-like positive reaction from the lower layer of the stratum granulosum to the upper layer of the stratum spinosum. CLD-2 ( F ) and CLD-5 ( G ) were positive in the salivary glands and the endothelium of blood vessel respectively, but were negative in the palatal epithelium. Background staining levels were checked by omitting the primary antibodies ( B : rabbit anti-goat IgG labeled with Alexa Fluor 488, E : goat anti-rabbit IgG labeled with Alexa Fluor 488). SC: stratum corneum, SB: stratum basale, Blue: nuclei stained with Hoechst. Bars = 25 μm ( A, B, C, D, E ), 50 μm ( F ), 10 μm ( G ).

    Techniques Used: Immunohistochemistry, Immunofluorescence, Fluorescence, Staining, Labeling

    31) Product Images from "The BAF (BRG1/BRM-Associated Factor) Chromatin-Remodeling Complex Exhibits Ethanol Sensitivity in Fetal Neural Progenitor Cells and Regulates Transcription at the Mir-9-2 Encoding Gene Locus"

    Article Title: The BAF (BRG1/BRM-Associated Factor) Chromatin-Remodeling Complex Exhibits Ethanol Sensitivity in Fetal Neural Progenitor Cells and Regulates Transcription at the Mir-9-2 Encoding Gene Locus

    Journal: Alcohol (Fayetteville, N.Y.)

    doi: 10.1016/j.alcohol.2017.01.003

    BRG1 protein expression in NSCs (a) Western immunoblot shows a single BRG1 band at ~238KD detected in the nuclear extract of neurosphere cultures but not in the cytoplasmic extract. (b) BRG1 protein levels in neurospheres after 0mg/dl, 120mg/dl and 320mg/dl ethanol treatment. Lower panel shows the corresponding immunoblot when probed for β-Actin as a loading control. (c) Bar graph, depicting the quantification of the western blot, shows that ethanol did not alter BRG1 protein expression, consistent with the lack of effect on BRG1 mRNA expression. The vertical axis shows the ratio of the band density of BRG1 to the ratio of the band density of β-Actin. Error bars indicates standard error of the mean. (d) Western blot analysis of BRG1 immunoprecipitation in cytoplasmic and nuclear cellular fractions probed with anti-BRG1 antibody. BRG1 is specifically precipitated with anti-BRG1 antibody (blue text), but not with an isotype-specific IgG control antibody (red text). Efficiency of anti-BRG1 immunoprecipitation is indicated by the relative depletion of BRG1 from the nuclear supernatant and enrichment in the immuno-precipitate (IP).
    Figure Legend Snippet: BRG1 protein expression in NSCs (a) Western immunoblot shows a single BRG1 band at ~238KD detected in the nuclear extract of neurosphere cultures but not in the cytoplasmic extract. (b) BRG1 protein levels in neurospheres after 0mg/dl, 120mg/dl and 320mg/dl ethanol treatment. Lower panel shows the corresponding immunoblot when probed for β-Actin as a loading control. (c) Bar graph, depicting the quantification of the western blot, shows that ethanol did not alter BRG1 protein expression, consistent with the lack of effect on BRG1 mRNA expression. The vertical axis shows the ratio of the band density of BRG1 to the ratio of the band density of β-Actin. Error bars indicates standard error of the mean. (d) Western blot analysis of BRG1 immunoprecipitation in cytoplasmic and nuclear cellular fractions probed with anti-BRG1 antibody. BRG1 is specifically precipitated with anti-BRG1 antibody (blue text), but not with an isotype-specific IgG control antibody (red text). Efficiency of anti-BRG1 immunoprecipitation is indicated by the relative depletion of BRG1 from the nuclear supernatant and enrichment in the immuno-precipitate (IP).

    Techniques Used: Expressing, Western Blot, Immunoprecipitation

    BRG1-containing complexes associate with both DNAse I-hypersensitive and insensitive sites on the miR-9-2 gene locus (a) Positional representation of POU5F1/Oct4, c-myc and REST transcription regulatory factor binding sites along the human pre-miR-9-2 gene locus identified with the UCSC genome browser ENCODE analysis hub track. Colored circles indicate locations for primer pairs for pri-miR-9-2 regions 1,2,3,4 and 5 and the pre-miR-9-2 coding region. (b) qPCR results of BRG1-ChIP from neurospheres. Primers were used to amplify 6 distinct positions along the pri-miR-9-2 gene locus and a genetically sparse region on mouse chromosome 6. Vertical axis shows fold change relative to an IgG pulldown control. Error bars indicate standard error of the mean. (c) qPCR of neurosphere DNA following digestion with 20, 60 and 120U of DNAse I at the 6 positions along the pri-miR-9-2 gene locus. The vertical axis shows fold amplification relative to DNAse I-untreated neurosphere DNA.
    Figure Legend Snippet: BRG1-containing complexes associate with both DNAse I-hypersensitive and insensitive sites on the miR-9-2 gene locus (a) Positional representation of POU5F1/Oct4, c-myc and REST transcription regulatory factor binding sites along the human pre-miR-9-2 gene locus identified with the UCSC genome browser ENCODE analysis hub track. Colored circles indicate locations for primer pairs for pri-miR-9-2 regions 1,2,3,4 and 5 and the pre-miR-9-2 coding region. (b) qPCR results of BRG1-ChIP from neurospheres. Primers were used to amplify 6 distinct positions along the pri-miR-9-2 gene locus and a genetically sparse region on mouse chromosome 6. Vertical axis shows fold change relative to an IgG pulldown control. Error bars indicate standard error of the mean. (c) qPCR of neurosphere DNA following digestion with 20, 60 and 120U of DNAse I at the 6 positions along the pri-miR-9-2 gene locus. The vertical axis shows fold amplification relative to DNAse I-untreated neurosphere DNA.

    Techniques Used: Binding Assay, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Amplification

    32) Product Images from "Anopheles Midgut Epithelium Evades Human Complement Activity by Capturing Factor H from the Blood Meal"

    Article Title: Anopheles Midgut Epithelium Evades Human Complement Activity by Capturing Factor H from the Blood Meal

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0003513

    Binding of mouse FH to A. stephensi mosquito proventriculus and anterior midgut epithelial surface in indirect immunofluorescence assays. Female A. stephensi mosquitoes were fed on mouse blood using the membrane glass feeder and were cut thereafter into sections. (A) An overlay image showing binding of mouse FH from the blood meal to the epithelial surface of the proventriculus (oval shaped region) and anterior midgut (narrow tube region) of the mosquito as a green fluorescence and DAPI nuclear staining as a blue fluorescence. (B) An overlay image of a consecutive mosquito section probed with Alexa Fluor 488 Donkey anti-goat IgG only to control for cross reactivity towards mosquito and blood proteins in the absence of anti-FH. Absence of green fluorescence in (B) indicated lack of cross reactivity. Scale bars are 50 μm.
    Figure Legend Snippet: Binding of mouse FH to A. stephensi mosquito proventriculus and anterior midgut epithelial surface in indirect immunofluorescence assays. Female A. stephensi mosquitoes were fed on mouse blood using the membrane glass feeder and were cut thereafter into sections. (A) An overlay image showing binding of mouse FH from the blood meal to the epithelial surface of the proventriculus (oval shaped region) and anterior midgut (narrow tube region) of the mosquito as a green fluorescence and DAPI nuclear staining as a blue fluorescence. (B) An overlay image of a consecutive mosquito section probed with Alexa Fluor 488 Donkey anti-goat IgG only to control for cross reactivity towards mosquito and blood proteins in the absence of anti-FH. Absence of green fluorescence in (B) indicated lack of cross reactivity. Scale bars are 50 μm.

    Techniques Used: Binding Assay, Immunofluorescence, Fluorescence, Staining

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    Immunocytochemistry:

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    Synthesized:

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    Cytometry:

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    Blocking Assay:

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    Enzyme-linked Immunosorbent Assay:

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    Incubation:

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    Mass Spectrometry:

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    Recombinase Polymerase Amplification:

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    High Performance Liquid Chromatography:

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    Flow Cytometry:

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    Concentration Assay:

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    Infection:

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    Article Title: Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG
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    Labeling:

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    Article Snippet: .. Materials and Methods Streptavidin–functionalized Microparticle (MP) (Dynabeads M-280 with a diameter of 2.8 μm), biotinylated polyclonal rabbit anti-goat IgG (rAb) and goat anti-rabbit IgG (goat IgG) (labeled with Alexa Fluor 488) were bought from Life Technologies (Carlsbad, CA, USA). ..

    Inhibition:

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    Recombinant:

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    Article Snippet: Protein and antigenic profiles of bacterial and recombinant protein preparations were analysed by Coomassie (Bio-Rad) and immunoblotting methods, respectively. .. Immunoblotting was performed using sera from mice experimentally infected with smooth SE-wt or from EDA hyperimmunized rabbits as primary antibodies and horseradish anti-mouse IgG or goat anti-rabbit IgG (ThermoFisher) as secondary antibodies, and the reaction was developed with diaminobenzidine.

    Immunofluorescence:

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    Article Title: Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG
    Article Snippet: The C. burnetii infection rate was determined at 4 h and at 5 days postinfection by an indirect immunofluorescence assay (IFA), as described previously ( ). .. Cells were fixed with 2% paraformaldehyde for 15 min and permeabilized with cold methanol for 10 min. Rabbit anti-NMI polyclonal antibodies (1:500) were used to stain intracellular C. burnetii , followed by incubation with goat anti-rabbit IgG (Invitrogen) (1:500).

    Fluorescence:

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    Mutagenesis:

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    Isolation:

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    Immunohistochemistry:

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    Microscopy:

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    Article Snippet: Antibodies were used at the following dilutions: anti-Pc (1:1000), anti-E(z) (1:1000), anti-Pho (1:1000), anti-Ubx (Developmental Studies Hybridoma Bank; 1:1000), anti-Mbf1 ( ; 1:500), goat anti-rabbit IgG or anti-mouse IgG Alexa488 (Molecular Probes, A72731 and A32723; 1:2000) and anti-rabbit IgG-Cy5 (Jackson ImmunoResearch, 111-225-144; 1:500). .. Images were acquired with an LSM510 META confocal microscope (Zeiss).

    Article Title: Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG
    Article Snippet: Cells were fixed with 2% paraformaldehyde for 15 min and permeabilized with cold methanol for 10 min. Rabbit anti-NMI polyclonal antibodies (1:500) were used to stain intracellular C. burnetii , followed by incubation with goat anti-rabbit IgG (Invitrogen) (1:500). .. Host nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI), and slides were examined by fluorescence microscopy.

    Purification:

    Article Title: Defects in lysosomal maturation facilitate the activation of innate sensors in systemic lupus erythematosus
    Article Snippet: Antibodies specific for LAMP1 and CD11b were purchased from BD Biosciences; LC3A from Cell Signaling; goat anti-rabbit IgG and rabbit anti-goat IgG from Molecular Probes; AIM2, ASC, TRIM21, p65, IRAK1, IRF3, and IRF7 from Santa Cruz Biotechnologies; anti-IgG from Jackson ImmunoResearch; and anti-LPS ( Escherichia coli J5) from Thermo Scientific. .. Antibodies specific to Smith (Sm; 2.12.3), nucleosome (PL2-3), CD16/32 (2.4G2), and TNP (Hy1.2) were purified from hybridoma culture supernatant by using protein G-Sepharose (GE Healthcare) then left unlabeled or conjugated with Alexa Fluor according to the manufacturer’s instructions (Molecular Probes).

    Article Title: Shadow Masking for Nanomaterial-Based Biosensors Incorporated with a Microfluidic Device
    Article Snippet: .. Goat anti-rabbit immunoglobulin G (IgG, 2 mg/mL, containing 0.1 M NaP, 0.1 M NaCl, and 5 mM azide, pH 7.5) antibodies (Abs) and IgG protein (purified rabbit IgG, 5.0 mg/mL, in 10 mM PBS, pH 7.4, containing 0.1% sodium azide) were obtained from Invitrogen. ..

    Selection:

    Article Title: IMMUNODOMINANT EPITOPE AND PROPERTIES OF PYROGLUTAMATE-MODIFIED A?-SPECIFIC ANTIBODIES PRODUCED IN RABBITS
    Article Snippet: Paragraph title: 2.6. Affinity selection of phages binding to anti- AβN3(pE) antibodies ... MaxiSorp microtiter plates were coated with goat anti-rabbit IgG (Zymed) at a concentration of 5 μg/ml 1 h at 37°C, washed and blocked with PBS-2%BSA.

    Immunostaining:

    Article Title: Mbf1 ensures Polycomb silencing by protecting E(z) mRNA from degradation by Pacman
    Article Snippet: Paragraph title: Immunostaining ... Antibodies were used at the following dilutions: anti-Pc (1:1000), anti-E(z) (1:1000), anti-Pho (1:1000), anti-Ubx (Developmental Studies Hybridoma Bank; 1:1000), anti-Mbf1 ( ; 1:500), goat anti-rabbit IgG or anti-mouse IgG Alexa488 (Molecular Probes, A72731 and A32723; 1:2000) and anti-rabbit IgG-Cy5 (Jackson ImmunoResearch, 111-225-144; 1:500).

    FACS:

    Article Title: Advantages of Papio anubis for preclinical testing of immunotoxicity of candidate therapeutic antagonist antibodies targeting CD28
    Article Snippet: FR104 staining was performed with a polyethylene glycol (PEG) rabbit mAb (Epitomics) followed by a fluorescent goat anti-rabbit IgG (Invitrogen). .. Samples were acquired on a BD FACS CANTOTM flow cytometer (BD Bioscience) and analyzed with FlowJo software.

    Mouse Assay:

    Article Title: The extradomain a of fibronectin enhances the efficacy of lipopolysaccharide defective Salmonella bacterins as vaccines in mice
    Article Snippet: .. Immunoblotting was performed using sera from mice experimentally infected with smooth SE-wt or from EDA hyperimmunized rabbits as primary antibodies and horseradish anti-mouse IgG or goat anti-rabbit IgG (ThermoFisher) as secondary antibodies, and the reaction was developed with diaminobenzidine. .. Proteins were quantified by the Bradford method (Bio-Rad).

    SDS Page:

    Article Title: DC3, the 21-kDa Subunit of the Outer Dynein Arm-Docking Complex (ODA-DC), Is a Novel EF-Hand Protein Important for Assembly of Both the Outer Arm and the ODA-DC
    Article Snippet: Paragraph title: SDS-PAGE and Western Blotting ... Goat anti-rabbit IgG coupled to horseradish peroxidase (HRP; Pierce, Rockford, IL) was used as the secondary antibody.

    Software:

    Article Title: Advantages of Papio anubis for preclinical testing of immunotoxicity of candidate therapeutic antagonist antibodies targeting CD28
    Article Snippet: FR104 staining was performed with a polyethylene glycol (PEG) rabbit mAb (Epitomics) followed by a fluorescent goat anti-rabbit IgG (Invitrogen). .. Samples were acquired on a BD FACS CANTOTM flow cytometer (BD Bioscience) and analyzed with FlowJo software.

    Article Title: DC3, the 21-kDa Subunit of the Outer Dynein Arm-Docking Complex (ODA-DC), Is a Novel EF-Hand Protein Important for Assembly of Both the Outer Arm and the ODA-DC
    Article Snippet: Goat anti-rabbit IgG coupled to horseradish peroxidase (HRP; Pierce, Rockford, IL) was used as the secondary antibody. .. The relative amount of protein in bands on western blots was quantitated using ImageQuant software (Amersham Biosciences, Piscataway, NJ); mutant values were calculated relative to wild type using either IC140 or RSP-1 as a loading control.

    Negative Control:

    Article Title: The extradomain a of fibronectin enhances the efficacy of lipopolysaccharide defective Salmonella bacterins as vaccines in mice
    Article Snippet: Unlabeled SE-wt bacterins were used as negative control. .. Immunoblotting was performed using sera from mice experimentally infected with smooth SE-wt or from EDA hyperimmunized rabbits as primary antibodies and horseradish anti-mouse IgG or goat anti-rabbit IgG (ThermoFisher) as secondary antibodies, and the reaction was developed with diaminobenzidine.

    Binding Assay:

    Article Title: IMMUNODOMINANT EPITOPE AND PROPERTIES OF PYROGLUTAMATE-MODIFIED A?-SPECIFIC ANTIBODIES PRODUCED IN RABBITS
    Article Snippet: Paragraph title: 2.6. Affinity selection of phages binding to anti- AβN3(pE) antibodies ... MaxiSorp microtiter plates were coated with goat anti-rabbit IgG (Zymed) at a concentration of 5 μg/ml 1 h at 37°C, washed and blocked with PBS-2%BSA.

    In Vitro:

    Article Title: Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG
    Article Snippet: Paragraph title: C. burnetii inhibition assay in vitro . ... Cells were fixed with 2% paraformaldehyde for 15 min and permeabilized with cold methanol for 10 min. Rabbit anti-NMI polyclonal antibodies (1:500) were used to stain intracellular C. burnetii , followed by incubation with goat anti-rabbit IgG (Invitrogen) (1:500).

    Immunoprecipitation:

    Article Title: Fibroblast Growth Factor Homologous Factor 2B: Association with Nav1.6 and Selective Colocalization at Nodes of Ranvier of Dorsal Root Axons
    Article Snippet: The mouse anti-IgG antibody used as a control for immunoprecipitation was purchased from Vector Laboratories (Burlingame, CA). .. Secondary antibodies 568 F(ab′)2 fragment of goat anti-rabbit IgG and Alexa Fluor 488 F(ab′)2 fragment of goat anti-rabbit IgG (Molecular Probes) were used in the colocalization experiments of FHF2 and Nav 1.6 in the hippocampus.

    Staining:

    Article Title: Control of somatic tissue differentiation by the long non-coding RNA TINCR
    Article Snippet: The secondary antibodies used were Alexa-555-conjugated goat anti-mouse and goat anti-rabbit IgG (Molecular Probes, 1:300 dilution), and Alexa-488-conjugated goat anti-rabbit and goat anti-mouse IgG (Molecular Probes, 1:300 dilution). .. For histological analysis, human organotypic epidermal tissue was fixed in 10% formalin (Sigma-Aldrich), embedded in paraffin, sectioned and stained with haematoxylin and eosin.

    Article Title: Advantages of Papio anubis for preclinical testing of immunotoxicity of candidate therapeutic antagonist antibodies targeting CD28
    Article Snippet: .. FR104 staining was performed with a polyethylene glycol (PEG) rabbit mAb (Epitomics) followed by a fluorescent goat anti-rabbit IgG (Invitrogen). .. CD28 receptor occupancy by FR104 on T lymphocytes was determined by performing the ratio of median fluorescent intensity (MFI) of FR104 staining between an unmodified blood sample and a blood sample incubated for 30 min at room temperature with a saturating concentration of FR104 (5 µg/ml).

    Article Title: Epstein–Barr Virus Lytic Reactivation Induces IgG4 Production by Host B Lymphocytes in Graves' Disease Patients and Controls: A Subset of Graves' Disease Is an IgG4-Related Disease-Like Condition
    Article Snippet: .. We used 1 μg/106 cells of an anti-human IgG4 rabbit monoclonal antibody, clone RM120 (Abnova, Taipei, Taiwan) to stain surface IgG4, and 1 μg/106 cells of goat anti-rabbit IgG (H + L) (Alexa Fluor® 647) (Invitrogen, Carlsbad, CA) as a secondary antibody. .. Fluorostained samples were analyzed and sorted with the cell sorter, MoFlo XDP (Beckman Coulter, Fullerton, CA).

    Article Title: Knockdown of lncRNA-ATB suppresses autocrine secretion of TGF-β2 by targeting ZNF217 via miR-200c in keloid fibroblasts
    Article Snippet: Paragraph title: Immunohistochemical staining, microscopy, and image analysis ... The primary antibody used was anti-ZNF217 rabbit polyclonal antibody (1:200; ab181741, Abcam, Cambridge, UK), and goat anti-rabbit IgG (Alexa Fluor 594, Invitrogen) was used as a secondary antibody.

    Article Title: Formalin-Inactivated Coxiella burnetii Phase I Vaccine-Induced Protection Depends on B Cells To Produce Protective IgM and IgG
    Article Snippet: .. Cells were fixed with 2% paraformaldehyde for 15 min and permeabilized with cold methanol for 10 min. Rabbit anti-NMI polyclonal antibodies (1:500) were used to stain intracellular C. burnetii , followed by incubation with goat anti-rabbit IgG (Invitrogen) (1:500). .. Host nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI), and slides were examined by fluorescence microscopy.

    Fluorescence In Situ Hybridization:

    Article Title: DC3, the 21-kDa Subunit of the Outer Dynein Arm-Docking Complex (ODA-DC), Is a Novel EF-Hand Protein Important for Assembly of Both the Outer Arm and the ODA-DC
    Article Snippet: Western blotting was performed according to standard procedures ( ) using 5% nonfat dry milk and 1% gelatin from cold water fish skin (Sigma, St. Louis, MO) in 10 mM Tris, pH 7.5, 166 mM NaCl, 0.05% Tween 20 as blocking agent. .. Goat anti-rabbit IgG coupled to horseradish peroxidase (HRP; Pierce, Rockford, IL) was used as the secondary antibody.

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    Thermo Scientific MagnaBind Beads provide a convenient method for magnetic separation of antibodies antigens lectins enzymes nucleic acids and cells using affinity binding To remove the MagnaBind Beads from the
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    Thermo Fisher rabbit anti goat igg hrp conjugated antibody
    The immunized mouse serum <t>IgG</t> (H+L) titer and IgG subtypes were detected using the indirect ELISA assay. (A) IgG titers of BALB/c mice immunized with rEF-Tu, rHSP70, Mo extracts and PBS sera of each groups were collected on days 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 DAI. (B) Determination of IgG subtypes in sera of the immunized mice. The sera of each group were collected at 35 DAI. ELISA plates were coated with purified rEF-Tu proteins or rHSP70 proteins or Mo extracts of M . ovipneumoniae wild strain Mo-1 at a concentration of 100 ng per well. Anti-mouse IgG (H+L) or IgG1 or IgG2a <t>HRP-conjugated</t> antibodies were used as secondary antibodies. Asterisks indicates the results of the One-Way ANOVA using the Tukey test, compared with the PBS negative control group, with P
    Rabbit Anti Goat Igg Hrp Conjugated Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 488 conjugated goat anti rabbit igg secondary antibody
    Determination of AvBD8 protein expression in immune cell lines by performing immunocytochemical analysis. Cultured chicken CU91 T-, DT40 B-, HD11 macrophage-, and OU2 fibroblast-cell lines were incubated with the rabbit anti-AvBD8 primary antibody, followed by incubation with the <t>Alexa</t> Fluor 488-conjugated goat anti-rabbit <t>IgG</t> secondary antibody, and were counterstained with 4′,6-diamidino-2-phenylindole. Control cells were incubated with secondary antibody only. AvBD8, avian beta-defensin 8; IgG, immunoglobulin G. Scale bar: 20 μm.
    Alexa Fluor 488 Conjugated Goat Anti Rabbit Igg Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher fitc conjugated goat anti rabbit igg
    Pf MSP8 is expressed during P. falciparum gametocyte development but is absent on the surface of activated macrogametes. (A) Detection of Pf MSP8 expression in fixed and permeabilized P. falciparum gametocytes (stages II to V) by an immunofluorescence assay with rabbit anti-r Pf MSP8 <t>IgG</t> or control IgG followed by <t>FITC-conjugated</t> secondary antibodies. DAPI was used to stain parasite DNA. (B) Analysis of Pf MSP8 expression on activated, live P. falciparum macrogametes by an immunofluorescence assay, as described above, with rabbit anti-r Pf MSP8 IgG. Samples were costained with MAb 4B7, which is specific for Pf s25 (MAb 4B7), followed by TRITC-conjugated secondary IgG. DIC, differential interference contrast.
    Fitc Conjugated Goat Anti Rabbit Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The immunized mouse serum IgG (H+L) titer and IgG subtypes were detected using the indirect ELISA assay. (A) IgG titers of BALB/c mice immunized with rEF-Tu, rHSP70, Mo extracts and PBS sera of each groups were collected on days 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 DAI. (B) Determination of IgG subtypes in sera of the immunized mice. The sera of each group were collected at 35 DAI. ELISA plates were coated with purified rEF-Tu proteins or rHSP70 proteins or Mo extracts of M . ovipneumoniae wild strain Mo-1 at a concentration of 100 ng per well. Anti-mouse IgG (H+L) or IgG1 or IgG2a HRP-conjugated antibodies were used as secondary antibodies. Asterisks indicates the results of the One-Way ANOVA using the Tukey test, compared with the PBS negative control group, with P

    Journal: PLoS ONE

    Article Title: Elongation Factor Tu and Heat Shock Protein 70 Are Membrane-Associated Proteins from Mycoplasma ovipneumoniae Capable of Inducing Strong Immune Response in Mice

    doi: 10.1371/journal.pone.0161170

    Figure Lengend Snippet: The immunized mouse serum IgG (H+L) titer and IgG subtypes were detected using the indirect ELISA assay. (A) IgG titers of BALB/c mice immunized with rEF-Tu, rHSP70, Mo extracts and PBS sera of each groups were collected on days 3, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 DAI. (B) Determination of IgG subtypes in sera of the immunized mice. The sera of each group were collected at 35 DAI. ELISA plates were coated with purified rEF-Tu proteins or rHSP70 proteins or Mo extracts of M . ovipneumoniae wild strain Mo-1 at a concentration of 100 ng per well. Anti-mouse IgG (H+L) or IgG1 or IgG2a HRP-conjugated antibodies were used as secondary antibodies. Asterisks indicates the results of the One-Way ANOVA using the Tukey test, compared with the PBS negative control group, with P

    Article Snippet: Following washing with PBST three times, membranes were incubated with rabbit anti-goat IgG HRP- conjugated antibody at 37°C for 1 h and exposed to SuperSignal® West Pico Chemiluminescent Substrate reagent for detection (Thermo Scientific, USA).

    Techniques: Indirect ELISA, Mouse Assay, Enzyme-linked Immunosorbent Assay, Purification, Concentration Assay, Negative Control

    Determination of AvBD8 protein expression in immune cell lines by performing immunocytochemical analysis. Cultured chicken CU91 T-, DT40 B-, HD11 macrophage-, and OU2 fibroblast-cell lines were incubated with the rabbit anti-AvBD8 primary antibody, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody, and were counterstained with 4′,6-diamidino-2-phenylindole. Control cells were incubated with secondary antibody only. AvBD8, avian beta-defensin 8; IgG, immunoglobulin G. Scale bar: 20 μm.

    Journal: Asian-Australasian Journal of Animal Sciences

    Article Title: Expression and regulation of avian beta-defensin 8 protein in immune tissues and cell lines of chickens

    doi: 10.5713/ajas.17.0836

    Figure Lengend Snippet: Determination of AvBD8 protein expression in immune cell lines by performing immunocytochemical analysis. Cultured chicken CU91 T-, DT40 B-, HD11 macrophage-, and OU2 fibroblast-cell lines were incubated with the rabbit anti-AvBD8 primary antibody, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody, and were counterstained with 4′,6-diamidino-2-phenylindole. Control cells were incubated with secondary antibody only. AvBD8, avian beta-defensin 8; IgG, immunoglobulin G. Scale bar: 20 μm.

    Article Snippet: After blocking, the cells were incubated overnight with the rabbit anti-AvBD8 primary antibody (dilution, 1:100) at 4°C, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody (dilution, 1:500) for 1 h. Control cells received secondary antibody only.

    Techniques: Expressing, Cell Culture, Incubation

    Determination of AvBD8 protein expression in immune tissues of male WL chickens by performing immunohistochemical analysis. Immunohistochemical analysis was performed to assess AvBD8 protein expression in the thymus, spleen, liver, small intestine, and ceca of male WL chickens aged 25 weeks. Frozen sections were incubated with the rabbit anti-AvBD8 primary antibody, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody, and were counterstained with 4′,6-diamidino-2-phenylindole. Control sections were incubated with secondary antibody only. Boxed region in the middle column is enlarged in the right column. AvBD8, avian beta-defensin 8; WL, White Leghorn; IgG, immunoglobulin G. Scale bar: 200 μm (left and middle columns) and 50 μm (right column). Arrowheads indicate strong AvBD8 signal in the intestinal mucosal layer.

    Journal: Asian-Australasian Journal of Animal Sciences

    Article Title: Expression and regulation of avian beta-defensin 8 protein in immune tissues and cell lines of chickens

    doi: 10.5713/ajas.17.0836

    Figure Lengend Snippet: Determination of AvBD8 protein expression in immune tissues of male WL chickens by performing immunohistochemical analysis. Immunohistochemical analysis was performed to assess AvBD8 protein expression in the thymus, spleen, liver, small intestine, and ceca of male WL chickens aged 25 weeks. Frozen sections were incubated with the rabbit anti-AvBD8 primary antibody, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody, and were counterstained with 4′,6-diamidino-2-phenylindole. Control sections were incubated with secondary antibody only. Boxed region in the middle column is enlarged in the right column. AvBD8, avian beta-defensin 8; WL, White Leghorn; IgG, immunoglobulin G. Scale bar: 200 μm (left and middle columns) and 50 μm (right column). Arrowheads indicate strong AvBD8 signal in the intestinal mucosal layer.

    Article Snippet: After blocking, the cells were incubated overnight with the rabbit anti-AvBD8 primary antibody (dilution, 1:100) at 4°C, followed by incubation with the Alexa Fluor 488-conjugated goat anti-rabbit IgG secondary antibody (dilution, 1:500) for 1 h. Control cells received secondary antibody only.

    Techniques: Expressing, Immunohistochemistry, Incubation

    Pf MSP8 is expressed during P. falciparum gametocyte development but is absent on the surface of activated macrogametes. (A) Detection of Pf MSP8 expression in fixed and permeabilized P. falciparum gametocytes (stages II to V) by an immunofluorescence assay with rabbit anti-r Pf MSP8 IgG or control IgG followed by FITC-conjugated secondary antibodies. DAPI was used to stain parasite DNA. (B) Analysis of Pf MSP8 expression on activated, live P. falciparum macrogametes by an immunofluorescence assay, as described above, with rabbit anti-r Pf MSP8 IgG. Samples were costained with MAb 4B7, which is specific for Pf s25 (MAb 4B7), followed by TRITC-conjugated secondary IgG. DIC, differential interference contrast.

    Journal: Infection and Immunity

    Article Title: Evaluation of a Plasmodium-Specific Carrier Protein To Enhance Production of Recombinant Pfs25, a Leading Transmission-Blocking Vaccine Candidate

    doi: 10.1128/IAI.00486-17

    Figure Lengend Snippet: Pf MSP8 is expressed during P. falciparum gametocyte development but is absent on the surface of activated macrogametes. (A) Detection of Pf MSP8 expression in fixed and permeabilized P. falciparum gametocytes (stages II to V) by an immunofluorescence assay with rabbit anti-r Pf MSP8 IgG or control IgG followed by FITC-conjugated secondary antibodies. DAPI was used to stain parasite DNA. (B) Analysis of Pf MSP8 expression on activated, live P. falciparum macrogametes by an immunofluorescence assay, as described above, with rabbit anti-r Pf MSP8 IgG. Samples were costained with MAb 4B7, which is specific for Pf s25 (MAb 4B7), followed by TRITC-conjugated secondary IgG. DIC, differential interference contrast.

    Article Snippet: Bound IgG was detected with FITC-conjugated goat anti-rabbit IgG, FITC-conjugated goat anti-mouse IgG, or tetramethylrhodamine isothiocyanate (TRITC)-labeled goat anti-mouse IgG as needed and then analyzed by fluorescence microscopy as described above.

    Techniques: Expressing, Immunofluorescence, Staining