novared  (Vector Laboratories)


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    Name:
    VECTOR NovaRED Peroxidase HRP Substrate Kit
    Description:
    Vector NovaRED Peroxidase Substrate Kit has greater sensitivity than conventional substratesConsistent and reliableIdeal for IHC ICC ISH and blotsPermanent mountingIdeal for single and multiple labelingStock solutions supplied in convenient dropper bottles promoting ease of handlingNo wait times for mixing and dissolving powders or tabletsOne year expiry dateSufficient reagents to produce 300 ml of working solution Vector NovaRED and ImmPACT NovaRED HRP substrates produce red reaction products Unlike AEC sections stained with either substrate should be dehydrated cleared and permanently mounted Both chromogens are useful as alternatives to DAB or as a second color for multiple antigen labeling Vector NovaRED and ImmPACT NovaRED substrates also provide excellent color contrast in pigmented tissue such as melanoma With the aid of imaging systems and software the spectral profile of both substrates can be distinguished from other enzyme substrates in applications where antigens are co localized Sections stained with either substrate also can be viewed by darkfield microscopy Vector NovaRED and ImmPACT NovaRED substrates can be used for both manual and automated staining methods Vector NovaRED Substrate Kit contains stock solutions in convenient dropper bottles The sensitivity of Vector NovaRED substrate is equivalent to Vector DAB Substrate and 4 times greater than AEC This product can also be used on blots This kit provides all of the necessary reagents to prepare about 300 ml of working solution When stored at 4 °C this kit is stable for one year
    Catalog Number:
    sk-4800
    Price:
    None
    Category:
    Protein chromogenic detection reagents or kits or substrates
    Size:
    1 Kit
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    Structured Review

    Vector Laboratories novared
    VECTOR NovaRED Peroxidase HRP Substrate Kit
    Vector NovaRED Peroxidase Substrate Kit has greater sensitivity than conventional substratesConsistent and reliableIdeal for IHC ICC ISH and blotsPermanent mountingIdeal for single and multiple labelingStock solutions supplied in convenient dropper bottles promoting ease of handlingNo wait times for mixing and dissolving powders or tabletsOne year expiry dateSufficient reagents to produce 300 ml of working solution Vector NovaRED and ImmPACT NovaRED HRP substrates produce red reaction products Unlike AEC sections stained with either substrate should be dehydrated cleared and permanently mounted Both chromogens are useful as alternatives to DAB or as a second color for multiple antigen labeling Vector NovaRED and ImmPACT NovaRED substrates also provide excellent color contrast in pigmented tissue such as melanoma With the aid of imaging systems and software the spectral profile of both substrates can be distinguished from other enzyme substrates in applications where antigens are co localized Sections stained with either substrate also can be viewed by darkfield microscopy Vector NovaRED and ImmPACT NovaRED substrates can be used for both manual and automated staining methods Vector NovaRED Substrate Kit contains stock solutions in convenient dropper bottles The sensitivity of Vector NovaRED substrate is equivalent to Vector DAB Substrate and 4 times greater than AEC This product can also be used on blots This kit provides all of the necessary reagents to prepare about 300 ml of working solution When stored at 4 °C this kit is stable for one year
    https://www.bioz.com/result/novared/product/Vector Laboratories
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    novared - by Bioz Stars, 2021-03
    97/100 stars

    Images

    1) Product Images from "Expression of C4.4A, a Structural uPAR Homolog, Reflects Squamous Epithelial Differentiation in the Adult Mouse and during Embryogenesis"

    Article Title: Expression of C4.4A, a Structural uPAR Homolog, Reflects Squamous Epithelial Differentiation in the Adult Mouse and during Embryogenesis

    Journal: Journal of Histochemistry and Cytochemistry

    doi: 10.1369/0022155410394859

    C4.4A expression in the mouse skin, eye, lung, and bladder. Paraffin-embedded tissue sections were incubated with our polyclonal rabbit anti-C4.4A antibody (2.5 µg/ml), stained with immunoperoxidase, and visualized by the NovaRed chromogene. Detection
    Figure Legend Snippet: C4.4A expression in the mouse skin, eye, lung, and bladder. Paraffin-embedded tissue sections were incubated with our polyclonal rabbit anti-C4.4A antibody (2.5 µg/ml), stained with immunoperoxidase, and visualized by the NovaRed chromogene. Detection

    Techniques Used: Expressing, Incubation, Staining

    2) Product Images from "Parecoxib is neuroprotective in spontaneously hypertensive rats after transient middle cerebral artery occlusion: a divided treatment response?"

    Article Title: Parecoxib is neuroprotective in spontaneously hypertensive rats after transient middle cerebral artery occlusion: a divided treatment response?

    Journal: Journal of Neuroinflammation

    doi: 10.1186/1742-2094-3-31

    COX-2 and NeuN double stains 24 hours and one week after tMCAo . The COX-2 IHC was developed with nickel-enhanced DAB (black), whereas NeuN was visualized with NovaRed ® (brownish red). The images 5A and 5B are obtained from a pilot study where the animal was euthanized 24 hours after tMCAo. 5A visualizes a relatively small neocortical infarct in the right hemisphere. The box delineates a part of the ischemic border zone that is shown at forty times magnification in 5B . The penumbra contains large swollen neurons that express the membrane-bound COX-2 enzyme. In the infarct core the neurons tend to be small and star-shaped due to irreversible neuronal death. 5C and 5E are from a saline-treated animal one week after tMCAo. Forty times magnifications of the boxes are shown in 5D and 5F . The neurons in the border zone on Day 8 after ischemic injury showed a perinuclear expression pattern of the COX-2 enzyme ( 5D ). COX-2 + neurons can be found in areas like the neocortex, piriform cortex and the DG of the hippocampus under normal conditions. 5F shows COX-2 expressed in dendrites of neurons in the molecular cell layer of the DG. The scale bar in 5A is 5 mm, whereas the scale bars in 5B , 5D and 5F equals 50 μm.
    Figure Legend Snippet: COX-2 and NeuN double stains 24 hours and one week after tMCAo . The COX-2 IHC was developed with nickel-enhanced DAB (black), whereas NeuN was visualized with NovaRed ® (brownish red). The images 5A and 5B are obtained from a pilot study where the animal was euthanized 24 hours after tMCAo. 5A visualizes a relatively small neocortical infarct in the right hemisphere. The box delineates a part of the ischemic border zone that is shown at forty times magnification in 5B . The penumbra contains large swollen neurons that express the membrane-bound COX-2 enzyme. In the infarct core the neurons tend to be small and star-shaped due to irreversible neuronal death. 5C and 5E are from a saline-treated animal one week after tMCAo. Forty times magnifications of the boxes are shown in 5D and 5F . The neurons in the border zone on Day 8 after ischemic injury showed a perinuclear expression pattern of the COX-2 enzyme ( 5D ). COX-2 + neurons can be found in areas like the neocortex, piriform cortex and the DG of the hippocampus under normal conditions. 5F shows COX-2 expressed in dendrites of neurons in the molecular cell layer of the DG. The scale bar in 5A is 5 mm, whereas the scale bars in 5B , 5D and 5F equals 50 μm.

    Techniques Used: Immunohistochemistry, Expressing

    BrdU, ED-1 and NeuN stains one week after tMCAo . 9A to 9F show the IHC for BrdU and ED-1. BrdU was developed with nickel-enhanced DAB (black), whereas the activated microglia marker ED-1 was visualized with NovaRed ® (brownish red). 9A shows a representative example of a tMCAo animal one week after surgery. Four and forty time magnifications of the boxes in 9A and 9B are shown in 9B and 9C , respectively. The BrdU + cells are typically found in clusters in the subgranular cell layer in DG. Our counting procedure started with delineating the DG. Hereafter, the BrdU + cells in the whole DG were counted at forty times magnification ( 9C ). We generally observed a very scarce ED-1 expression in the DG unless the hippocampus was directly affected by ischemic injury. 9D to 9I show an example of ischemia affecting the right hippocampus. The boxed area in 9D is shown at higher magnification in 9E . Activated microglia was abundantly seen in a part of the CA-1 and the whole DG. 9F shows a forty time magnification of the box in 9E . Clearly, activated microglia had an intimate relation to an increasing number of BrdU + cells. Panel 9G to 9I show NeuN IHC developed with nickel-enhanced DAB. 9G and 9H correspond to 9D and 9E . The boxed area in 9H of the CA-1 is shown at forty time magnification in 9I . Note the ischemic degeneration of this part of the CA-1. The scale bar in 9A is 5 mm. In 9B the scale bar represents 300 mm, and in 9E and 9H 1 mm. The scale bars in 9C , 9F and 9I equals 50 μm.
    Figure Legend Snippet: BrdU, ED-1 and NeuN stains one week after tMCAo . 9A to 9F show the IHC for BrdU and ED-1. BrdU was developed with nickel-enhanced DAB (black), whereas the activated microglia marker ED-1 was visualized with NovaRed ® (brownish red). 9A shows a representative example of a tMCAo animal one week after surgery. Four and forty time magnifications of the boxes in 9A and 9B are shown in 9B and 9C , respectively. The BrdU + cells are typically found in clusters in the subgranular cell layer in DG. Our counting procedure started with delineating the DG. Hereafter, the BrdU + cells in the whole DG were counted at forty times magnification ( 9C ). We generally observed a very scarce ED-1 expression in the DG unless the hippocampus was directly affected by ischemic injury. 9D to 9I show an example of ischemia affecting the right hippocampus. The boxed area in 9D is shown at higher magnification in 9E . Activated microglia was abundantly seen in a part of the CA-1 and the whole DG. 9F shows a forty time magnification of the box in 9E . Clearly, activated microglia had an intimate relation to an increasing number of BrdU + cells. Panel 9G to 9I show NeuN IHC developed with nickel-enhanced DAB. 9G and 9H correspond to 9D and 9E . The boxed area in 9H of the CA-1 is shown at forty time magnification in 9I . Note the ischemic degeneration of this part of the CA-1. The scale bar in 9A is 5 mm. In 9B the scale bar represents 300 mm, and in 9E and 9H 1 mm. The scale bars in 9C , 9F and 9I equals 50 μm.

    Techniques Used: Immunohistochemistry, Marker, Expressing

    3) Product Images from "Histopathological Growth Pattern, Proteolysis and Angiogenesis in Chemonaive Patients Resected for Multiple Colorectal Liver Metastases"

    Article Title: Histopathological Growth Pattern, Proteolysis and Angiogenesis in Chemonaive Patients Resected for Multiple Colorectal Liver Metastases

    Journal: Journal of Oncology

    doi: 10.1155/2012/907971

    Localisation of uPAR and CD68-positive macrophages in colorectal liver metastases. Adjacent sections from a liver metastasis with desmoplastic growth (a)–(c), a liver metastasis with pushing growth pattern (d)–(f), and a liver metastasis with replacement growth pattern (g)–(i) were stained for uPAR (a), (b), (d), (e), (g), (h) or double stained for cytokeratin and CD68 (c), (f), (i). The uPAR-immunoreactivity was visualised by NovaRed, cytokeratin with Permanent Red, and CD68 with DAB. In the liver metastasis with desmoplastic growth pattern, strong expression of uPAR is seen in macrophages located at the invasive front within the desmoplastic zone (a), (b). Large numbers of macrophages were found at the front of the metastasis (c). In the liver metastasis with pushing growth pattern, uPAR expression is seen in a few macrophages and fibroblast-like cells intermingled with the tumour cells (d), (e). Few macrophages are located at the metastasis/liver parenchyma interface (f). In the liver metastasis with replacement growth pattern, uPAR expression is confined to some macrophages and fibroblast-like cells located between the tumour cells (g)-(h). Few macrophages are located at the metastasis/liver parenchyma interface (i). Black arrows points at the tumour periphery. Black arrowhead in (e) and (g) points at a uPAR-positive neutrophil, which is an internal positive control. Bar a: 100 μ m. Bar b: 50 μ m.
    Figure Legend Snippet: Localisation of uPAR and CD68-positive macrophages in colorectal liver metastases. Adjacent sections from a liver metastasis with desmoplastic growth (a)–(c), a liver metastasis with pushing growth pattern (d)–(f), and a liver metastasis with replacement growth pattern (g)–(i) were stained for uPAR (a), (b), (d), (e), (g), (h) or double stained for cytokeratin and CD68 (c), (f), (i). The uPAR-immunoreactivity was visualised by NovaRed, cytokeratin with Permanent Red, and CD68 with DAB. In the liver metastasis with desmoplastic growth pattern, strong expression of uPAR is seen in macrophages located at the invasive front within the desmoplastic zone (a), (b). Large numbers of macrophages were found at the front of the metastasis (c). In the liver metastasis with pushing growth pattern, uPAR expression is seen in a few macrophages and fibroblast-like cells intermingled with the tumour cells (d), (e). Few macrophages are located at the metastasis/liver parenchyma interface (f). In the liver metastasis with replacement growth pattern, uPAR expression is confined to some macrophages and fibroblast-like cells located between the tumour cells (g)-(h). Few macrophages are located at the metastasis/liver parenchyma interface (i). Black arrows points at the tumour periphery. Black arrowhead in (e) and (g) points at a uPAR-positive neutrophil, which is an internal positive control. Bar a: 100 μ m. Bar b: 50 μ m.

    Techniques Used: Staining, Expressing, Positive Control

    4) Product Images from "Histamine Induces Alzheimer's Disease-Like Blood Brain Barrier Breach and Local Cellular Responses in Mouse Brain Organotypic Cultures"

    Article Title: Histamine Induces Alzheimer's Disease-Like Blood Brain Barrier Breach and Local Cellular Responses in Mouse Brain Organotypic Cultures

    Journal: BioMed Research International

    doi: 10.1155/2015/937148

    Histamine leads to astrocyte activation in MBO cultures. (a) Control MBO cultures show few astrocytes that are GFAP-positive (visualized by NovaRED and indicated by arrows). (b) Histamine-treated MBO cultures show many intensely GFAP-positive astrocytes (visualized by NovaRED and indicated by arrows). In both (a) and (b), sections are from the middle portion of the MBO tissue (350–700 μ m from the slice surface) and counterstained with hematoxylin to show nucleus in purple as described in Section 2 . Scale bar, 100 μ m. (c) Quantification shows more GFAP-positive astrocytes in the histamine-treated MBO cultures compared to the controls. GFAP-positive astrocytes are counted from multiple images, normalized by the total cell numbers and plotted. Mean ± SEM is used to represent the variations within each group. ∗ P
    Figure Legend Snippet: Histamine leads to astrocyte activation in MBO cultures. (a) Control MBO cultures show few astrocytes that are GFAP-positive (visualized by NovaRED and indicated by arrows). (b) Histamine-treated MBO cultures show many intensely GFAP-positive astrocytes (visualized by NovaRED and indicated by arrows). In both (a) and (b), sections are from the middle portion of the MBO tissue (350–700 μ m from the slice surface) and counterstained with hematoxylin to show nucleus in purple as described in Section 2 . Scale bar, 100 μ m. (c) Quantification shows more GFAP-positive astrocytes in the histamine-treated MBO cultures compared to the controls. GFAP-positive astrocytes are counted from multiple images, normalized by the total cell numbers and plotted. Mean ± SEM is used to represent the variations within each group. ∗ P

    Techniques Used: Activation Assay

    Related Articles

    Immunohistochemistry:

    Article Title: 12E2
    Article Snippet: 12E2 cells were analyzed using BD Falcon culture slides coated with type IV collagen (Sigma) or 5-μm cryostat sections of 12E2-microinjected footpads. .. Immunohistochemistry was performed using a three-step biotin avidin horseradish peroxidase (HRP) (ABC) system with NovaRed substrate kit (Vector Labs, Burlingame, CA). .. Briefly, cells or sections were fixed in acetone at −20°C and then incubated with primary monoclonal antibody or isotype-matched control antibody for 1 hour at room temperature (RT).

    Avidin-Biotin Assay:

    Article Title: 12E2
    Article Snippet: 12E2 cells were analyzed using BD Falcon culture slides coated with type IV collagen (Sigma) or 5-μm cryostat sections of 12E2-microinjected footpads. .. Immunohistochemistry was performed using a three-step biotin avidin horseradish peroxidase (HRP) (ABC) system with NovaRed substrate kit (Vector Labs, Burlingame, CA). .. Briefly, cells or sections were fixed in acetone at −20°C and then incubated with primary monoclonal antibody or isotype-matched control antibody for 1 hour at room temperature (RT).

    Plasmid Preparation:

    Article Title: 12E2
    Article Snippet: 12E2 cells were collected by trypsinization 24 hours after CSFE labeling. .. CSFE-labeled 12E2 cells were detected in cryosections of footpads or cytospin preparations using polyclonal goat anti-fluorescein antibody (Vector) followed by horse anti-goat HRP (Vector) and development with NovaRed substrate kit (Vector). .. Single-cell suspensions of 12E2 or 3T3 (2 × 106 cells in 50 μl HBSS) were injected intradermally per footpad of syngeneic BALB/c AnCr ( n = 8), SCID/NCr (BALB/c background; n = 4) or allogeneic CBA/NCr ( n = 6) mice.

    Article Title: Receptor cleavage and P-selectin-dependent reduction of leukocyte adhesion in the spontaneously hypertensive rat
    Article Snippet: Primary antibodies targeting the extracellular domain (Santa Cruz Biotechnology, Santa Cruz, CA, USA; sc-10174; 12 μg/ml final concentration) or intracellular domain (Santa Cruz Biotechnology; sc-10176; 12 μg/ml final concentration) of PSGL-1 were applied for 90 min. For negative control, normal goat IgG (Santa Cruz Biotechnology; # sc-2028; 12 μg/ml final concentration) was applied in place of the primary antibody. .. Following primary antibody, secondary antibody conjugated to peroxidase (Vector Laboratories; #MP-7405; ImPRESS reagent anti-goat Ig peroxidase kit) was applied to the blood smears for 30 min. Lastly, Vector NovaRED, which is a substrate for peroxidase, was applied for 5 min (Vector Laboratories; #SK-4800), and a color change in red served as the reporter for antibody binding to PSGL-1. ..

    Staining:

    Article Title: Neuroanatomical Study of the A11 Diencephalospinal Pathway in the Non-Human Primate
    Article Snippet: After FG neuron revelation, sections were first incubated with an avidin/biotin blocking kit (Vector Laboratories) and then re-incubated with TH antibody (1∶10000) for one night at room temperature. .. TH-IR was revealed with the NovaRed substrate kit for peroxidase (Vector Laboratories) that stained the TH-positive neurons red. ..

    Article Title: Complete Remission of Mouse Melanoma after Temporally Fractionated Microbeam Radiotherapy
    Article Snippet: .. In addition, the slides were stained with the NovaRED substrate kit for peroxidase Reagent (MP-7451, Vector Labs, Burlingame, CA, USA) instead of DAB. ..

    Binding Assay:

    Article Title: Receptor cleavage and P-selectin-dependent reduction of leukocyte adhesion in the spontaneously hypertensive rat
    Article Snippet: Primary antibodies targeting the extracellular domain (Santa Cruz Biotechnology, Santa Cruz, CA, USA; sc-10174; 12 μg/ml final concentration) or intracellular domain (Santa Cruz Biotechnology; sc-10176; 12 μg/ml final concentration) of PSGL-1 were applied for 90 min. For negative control, normal goat IgG (Santa Cruz Biotechnology; # sc-2028; 12 μg/ml final concentration) was applied in place of the primary antibody. .. Following primary antibody, secondary antibody conjugated to peroxidase (Vector Laboratories; #MP-7405; ImPRESS reagent anti-goat Ig peroxidase kit) was applied to the blood smears for 30 min. Lastly, Vector NovaRED, which is a substrate for peroxidase, was applied for 5 min (Vector Laboratories; #SK-4800), and a color change in red served as the reporter for antibody binding to PSGL-1. ..

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    Vector Laboratories novared substrate kit
    Phenotypic and immunochemical characterization of 12E2 cells in vitro . A: By phase contrast microscopy, cells with numerous, thin, elongated dendrites characterized subconfluent (depicted here) and confluent cultures. B: Immunohistochemistry for the integral membrane protein (DEC-205) defined by NLDC-145 monoclonal antibody permitted further confirmation of the prominent membranous dendritic extensions of cultured 12E2 cells. C and D: Strong immunoreactivity for FXIIIa ( C ) and VCAM-1 ( D ) was restricted primarily to cell bodies, but not dendrites, of 12E2 cells. E and F: Western blot analyses of 12E2 lysates were positive for FXIIIa ( E ) and VCAM-1 ( F ). The positive band for FXIIIa was detected at approximately 80 kd and for VCAM-1 at approximately 100 kd. The molecular weight for 12E2-derived FXIIIa approximated that of purified human FXIII ( E The molecular weight for 12E2-derived VCAM-1 closely approximated that identified from VCAM-1+ lysates of the SV40-transformed murine endothelial cell line that constitutively expresses VCAM-1 ( F ). Original magnification: ×400 ( A to D ); <t>NovaRed</t> chromagen used in B to D .
    Novared Substrate Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/novared substrate kit/product/Vector Laboratories
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    novared substrate kit - by Bioz Stars, 2021-03
    97/100 stars
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    Phenotypic and immunochemical characterization of 12E2 cells in vitro . A: By phase contrast microscopy, cells with numerous, thin, elongated dendrites characterized subconfluent (depicted here) and confluent cultures. B: Immunohistochemistry for the integral membrane protein (DEC-205) defined by NLDC-145 monoclonal antibody permitted further confirmation of the prominent membranous dendritic extensions of cultured 12E2 cells. C and D: Strong immunoreactivity for FXIIIa ( C ) and VCAM-1 ( D ) was restricted primarily to cell bodies, but not dendrites, of 12E2 cells. E and F: Western blot analyses of 12E2 lysates were positive for FXIIIa ( E ) and VCAM-1 ( F ). The positive band for FXIIIa was detected at approximately 80 kd and for VCAM-1 at approximately 100 kd. The molecular weight for 12E2-derived FXIIIa approximated that of purified human FXIII ( E The molecular weight for 12E2-derived VCAM-1 closely approximated that identified from VCAM-1+ lysates of the SV40-transformed murine endothelial cell line that constitutively expresses VCAM-1 ( F ). Original magnification: ×400 ( A to D ); NovaRed chromagen used in B to D .

    Journal: The American Journal of Pathology

    Article Title: 12E2

    doi:

    Figure Lengend Snippet: Phenotypic and immunochemical characterization of 12E2 cells in vitro . A: By phase contrast microscopy, cells with numerous, thin, elongated dendrites characterized subconfluent (depicted here) and confluent cultures. B: Immunohistochemistry for the integral membrane protein (DEC-205) defined by NLDC-145 monoclonal antibody permitted further confirmation of the prominent membranous dendritic extensions of cultured 12E2 cells. C and D: Strong immunoreactivity for FXIIIa ( C ) and VCAM-1 ( D ) was restricted primarily to cell bodies, but not dendrites, of 12E2 cells. E and F: Western blot analyses of 12E2 lysates were positive for FXIIIa ( E ) and VCAM-1 ( F ). The positive band for FXIIIa was detected at approximately 80 kd and for VCAM-1 at approximately 100 kd. The molecular weight for 12E2-derived FXIIIa approximated that of purified human FXIII ( E The molecular weight for 12E2-derived VCAM-1 closely approximated that identified from VCAM-1+ lysates of the SV40-transformed murine endothelial cell line that constitutively expresses VCAM-1 ( F ). Original magnification: ×400 ( A to D ); NovaRed chromagen used in B to D .

    Article Snippet: CSFE-labeled 12E2 cells were detected in cryosections of footpads or cytospin preparations using polyclonal goat anti-fluorescein antibody (Vector) followed by horse anti-goat HRP (Vector) and development with NovaRed substrate kit (Vector).

    Techniques: In Vitro, Microscopy, Immunohistochemistry, Cell Culture, Western Blot, Molecular Weight, Derivative Assay, Purification, Transformation Assay

    Immunophenotyping and growth characteristics of 12E2 cells in footpad microinjection model at 1 week. A and B: Microinjection of control medium containing inert carbon failed to influence number or distribution of constitutive FXIIIaDD. C , D , and D inset : In contrast, syngeneic footpads microinjected with 12E2 cells produced a lesion composed of closely-aggregated FXIIIaDD. E: CFSE prelabeling of 12E2 cells before injection and detected immunohistochemically demonstrated that the experimentally introduced 12E2 cells, and not recipient (constitutive) FXIIIaDD, accounted for this lesion. CFSE label was of variable intensity within 12E2 cells, consistent with dilution consequent to proliferation ( E , inset ). F: The in situ growth pattern of 12E2 cells was characterized by interstitial permeation between collagen and smooth muscle fibers by conventional H E staining. G and H: Immunohistochemistry (here for CFSE) permitted detection of envelopment of dermal blood vessels ( G ) and nerves ( H ) by infiltrating 12E2 cells. This pattern differed from that of microinjected 3T3 fibroblasts, which failed to exhibit infiltrative growth. Similar findings were documented in footpads from SCID and allogeneic (CBA) mice, although the latter was associated with a brisk mononuclear cell infiltrate. Arrows indicate particulate carbon deposits ( A and B ). m, smooth muscle fiber ( F ); v, vessel ( G ); n, nerve fiber ( H ). Original magnifications: ×100 ( A , C , and E ); ×200 ( B and D ); ×400 ( D ( inset ), E and inset , and F to H ). Chromagen is NovaRed except in F , which is H E.

    Journal: The American Journal of Pathology

    Article Title: 12E2

    doi:

    Figure Lengend Snippet: Immunophenotyping and growth characteristics of 12E2 cells in footpad microinjection model at 1 week. A and B: Microinjection of control medium containing inert carbon failed to influence number or distribution of constitutive FXIIIaDD. C , D , and D inset : In contrast, syngeneic footpads microinjected with 12E2 cells produced a lesion composed of closely-aggregated FXIIIaDD. E: CFSE prelabeling of 12E2 cells before injection and detected immunohistochemically demonstrated that the experimentally introduced 12E2 cells, and not recipient (constitutive) FXIIIaDD, accounted for this lesion. CFSE label was of variable intensity within 12E2 cells, consistent with dilution consequent to proliferation ( E , inset ). F: The in situ growth pattern of 12E2 cells was characterized by interstitial permeation between collagen and smooth muscle fibers by conventional H E staining. G and H: Immunohistochemistry (here for CFSE) permitted detection of envelopment of dermal blood vessels ( G ) and nerves ( H ) by infiltrating 12E2 cells. This pattern differed from that of microinjected 3T3 fibroblasts, which failed to exhibit infiltrative growth. Similar findings were documented in footpads from SCID and allogeneic (CBA) mice, although the latter was associated with a brisk mononuclear cell infiltrate. Arrows indicate particulate carbon deposits ( A and B ). m, smooth muscle fiber ( F ); v, vessel ( G ); n, nerve fiber ( H ). Original magnifications: ×100 ( A , C , and E ); ×200 ( B and D ); ×400 ( D ( inset ), E and inset , and F to H ). Chromagen is NovaRed except in F , which is H E.

    Article Snippet: CSFE-labeled 12E2 cells were detected in cryosections of footpads or cytospin preparations using polyclonal goat anti-fluorescein antibody (Vector) followed by horse anti-goat HRP (Vector) and development with NovaRed substrate kit (Vector).

    Techniques: Produced, Injection, In Situ, Staining, Immunohistochemistry, Crocin Bleaching Assay, Mouse Assay

    Comparison of lesions produced by 12E2 microinjection model and human Kaposi’s sarcoma. A and B: Conventional histology of 12E2 microinjection sites after 1 week, showing infiltration of superficial ( A ) and deeper ( B ) dermal layers by bland spindle cells associated with dilated blood vessels. C to F: Immunohistochemistry of adjacent sections of 12E2 microinjection site (1 week) stained for FXIIIa ( C and E ) and the endothelial marker, CD31 ( D and F ), documenting angiogenesis associated with infiltration of 12E2 cells. G and H: Conventional histology of characteristic human KS lesion. I to L: Immunohistochemistry of adjacent sections of typical human KS lesion stained for FXIIIa ( I and K ) and CD31 ( J and L ). Note histological and immunohistochemical similarities to the 12E2 microinjection sites ( A to F ). *Corresponding adjacent foci in C to F , I and J , K , and L FXIIIa/CD31 pairs. Original magnifications: ×400 ( A , E to G , K , and L ) and ×200 ( B to D , H to J ). H E staining in A , B , G and H . NovaRed chromagen in C to F and I to L .

    Journal: The American Journal of Pathology

    Article Title: 12E2

    doi:

    Figure Lengend Snippet: Comparison of lesions produced by 12E2 microinjection model and human Kaposi’s sarcoma. A and B: Conventional histology of 12E2 microinjection sites after 1 week, showing infiltration of superficial ( A ) and deeper ( B ) dermal layers by bland spindle cells associated with dilated blood vessels. C to F: Immunohistochemistry of adjacent sections of 12E2 microinjection site (1 week) stained for FXIIIa ( C and E ) and the endothelial marker, CD31 ( D and F ), documenting angiogenesis associated with infiltration of 12E2 cells. G and H: Conventional histology of characteristic human KS lesion. I to L: Immunohistochemistry of adjacent sections of typical human KS lesion stained for FXIIIa ( I and K ) and CD31 ( J and L ). Note histological and immunohistochemical similarities to the 12E2 microinjection sites ( A to F ). *Corresponding adjacent foci in C to F , I and J , K , and L FXIIIa/CD31 pairs. Original magnifications: ×400 ( A , E to G , K , and L ) and ×200 ( B to D , H to J ). H E staining in A , B , G and H . NovaRed chromagen in C to F and I to L .

    Article Snippet: CSFE-labeled 12E2 cells were detected in cryosections of footpads or cytospin preparations using polyclonal goat anti-fluorescein antibody (Vector) followed by horse anti-goat HRP (Vector) and development with NovaRed substrate kit (Vector).

    Techniques: Produced, Immunohistochemistry, Staining, Marker

    Recombinant IFN-γ-induced Ia expression and allostimulatory activity by 12E2 cells in vitro . A to C: 12E2 cells were negative to weakly positive by immunohistochemistry for Ia ( A ), whereas exposure to increasing concentrations of recombinant IFN-γ induced progressively intense immunoreactivity ( B and C ). D: Fixed numbers of naïve B10.BR T cells were combined with decreasing numbers of the respective stimulator cell type (BALB/c-positive control splenocytes (□), unstimulated 12E2 cells (○), 12E2 cells with IFN-γ pretreatment (▪), and IFN-γ-pretreated 3T3-negative control murine fibroblasts (•) to attain the indicated stimulator:responder ratios. Proliferation was measured on day 4 of co-culture and the stimulation index (SI) was calculated (see Materials and Methods). Optimal BALB/c allogeneic control stimulation at higher stimulator:responder cell ratios is shown. However, correlative data could not be generated for 12E2 cells as a result of cell aggregation at these higher ratios. Original magnification, ×400 with NovaRed chromagen ( A to C ).

    Journal: The American Journal of Pathology

    Article Title: 12E2

    doi:

    Figure Lengend Snippet: Recombinant IFN-γ-induced Ia expression and allostimulatory activity by 12E2 cells in vitro . A to C: 12E2 cells were negative to weakly positive by immunohistochemistry for Ia ( A ), whereas exposure to increasing concentrations of recombinant IFN-γ induced progressively intense immunoreactivity ( B and C ). D: Fixed numbers of naïve B10.BR T cells were combined with decreasing numbers of the respective stimulator cell type (BALB/c-positive control splenocytes (□), unstimulated 12E2 cells (○), 12E2 cells with IFN-γ pretreatment (▪), and IFN-γ-pretreated 3T3-negative control murine fibroblasts (•) to attain the indicated stimulator:responder ratios. Proliferation was measured on day 4 of co-culture and the stimulation index (SI) was calculated (see Materials and Methods). Optimal BALB/c allogeneic control stimulation at higher stimulator:responder cell ratios is shown. However, correlative data could not be generated for 12E2 cells as a result of cell aggregation at these higher ratios. Original magnification, ×400 with NovaRed chromagen ( A to C ).

    Article Snippet: CSFE-labeled 12E2 cells were detected in cryosections of footpads or cytospin preparations using polyclonal goat anti-fluorescein antibody (Vector) followed by horse anti-goat HRP (Vector) and development with NovaRed substrate kit (Vector).

    Techniques: Recombinant, IA, Expressing, Activity Assay, In Vitro, Immunohistochemistry, Positive Control, Negative Control, Co-Culture Assay, Generated

    Development of sex cord-stromal tumors in TGFBR1-CA G9Cre mice. a-f Histological analysis of 8-week-old control and TGFBR1-CA G9Cre mice. Note the presence of hemorrhagic cysts ( c ; blue arrows) and hemorrhage ( d ; blue arrows) and neoplastic regions containing mitotic figures ( e and f ; red arrows) in TGFBR1-CA G9Cre ovaries compared with control ovaries ( a and b ). Panel f is a higher magnification image for the boxed region in panel ( e ). g and h Immunohistochemical analysis of Ki67 using 8-week-old TGFBR1-CA G9Cre ( g ) and control ( h ) ovaries. Experiment was performed using ABC method, and signals were developed using NovaRED Peroxidase Substrate Kit. Sections were counterstained with hematoxylin. Scale bar is representatively depicted in ( a ) and equals 12.5 μm ( f ), 25 μm ( e , g , and h ), 50 μm ( b ), and 250 μm ( a , c , and d ). H E staining and immunohistochemistry were conducted using 3-5 independent samples per group. i Gross ovarian tumor morphology of a 7-month-old mouse. Yellow arrows denote the ovary and ovarian tumors in the control and TGFBR1-CA G9Cre mice, respectively

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Disruption of postnatal folliculogenesis and development of ovarian tumor in a mouse model with aberrant transforming growth factor beta signaling

    doi: 10.1186/s12958-017-0312-z

    Figure Lengend Snippet: Development of sex cord-stromal tumors in TGFBR1-CA G9Cre mice. a-f Histological analysis of 8-week-old control and TGFBR1-CA G9Cre mice. Note the presence of hemorrhagic cysts ( c ; blue arrows) and hemorrhage ( d ; blue arrows) and neoplastic regions containing mitotic figures ( e and f ; red arrows) in TGFBR1-CA G9Cre ovaries compared with control ovaries ( a and b ). Panel f is a higher magnification image for the boxed region in panel ( e ). g and h Immunohistochemical analysis of Ki67 using 8-week-old TGFBR1-CA G9Cre ( g ) and control ( h ) ovaries. Experiment was performed using ABC method, and signals were developed using NovaRED Peroxidase Substrate Kit. Sections were counterstained with hematoxylin. Scale bar is representatively depicted in ( a ) and equals 12.5 μm ( f ), 25 μm ( e , g , and h ), 50 μm ( b ), and 250 μm ( a , c , and d ). H E staining and immunohistochemistry were conducted using 3-5 independent samples per group. i Gross ovarian tumor morphology of a 7-month-old mouse. Yellow arrows denote the ovary and ovarian tumors in the control and TGFBR1-CA G9Cre mice, respectively

    Article Snippet: Signals were developed using NovaRED Peroxidase Substrate Kit (Vector Laboratories).

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    Immunohistochemical analysis of ovarian tumor markers. a-j Expression of granulosa cell and germ cell markers in 8-week-old control and TGFBR1-CA G9Cre ovaries. Representative images from immunohistochemical analysis of FOXL2 ( a and b ), INHA ( c and d ), FOXO1 ( e and f ), AMH ( g and h ), and DDX4 ( i and j ) are shown. k and l Negative controls using isotype-matched rabbit and goat IgGs. Experiment was performed using ABC method, and signals were developed using NovaRED Peroxidase Substrate Kit. Sections were counterstained with hematoxylin. Five independent samples per group were utilized in this analysis. Scale bar is representatively depicted in ( a ) and equals 50 μm ( a - l )

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Disruption of postnatal folliculogenesis and development of ovarian tumor in a mouse model with aberrant transforming growth factor beta signaling

    doi: 10.1186/s12958-017-0312-z

    Figure Lengend Snippet: Immunohistochemical analysis of ovarian tumor markers. a-j Expression of granulosa cell and germ cell markers in 8-week-old control and TGFBR1-CA G9Cre ovaries. Representative images from immunohistochemical analysis of FOXL2 ( a and b ), INHA ( c and d ), FOXO1 ( e and f ), AMH ( g and h ), and DDX4 ( i and j ) are shown. k and l Negative controls using isotype-matched rabbit and goat IgGs. Experiment was performed using ABC method, and signals were developed using NovaRED Peroxidase Substrate Kit. Sections were counterstained with hematoxylin. Five independent samples per group were utilized in this analysis. Scale bar is representatively depicted in ( a ) and equals 50 μm ( a - l )

    Article Snippet: Signals were developed using NovaRED Peroxidase Substrate Kit (Vector Laboratories).

    Techniques: Immunohistochemistry, Expressing

    Whole mount mesentery microvessels labeled with extracellular P-selectin antibody using Vector NovaRED as reporter substrate.

    Journal: Journal of Leukocyte Biology

    Article Title: Receptor cleavage and P-selectin-dependent reduction of leukocyte adhesion in the spontaneously hypertensive rat

    doi: 10.1189/jlb.0112010

    Figure Lengend Snippet: Whole mount mesentery microvessels labeled with extracellular P-selectin antibody using Vector NovaRED as reporter substrate.

    Article Snippet: Following primary antibody, secondary antibody conjugated to peroxidase (Vector Laboratories; #MP-7405; ImPRESS reagent anti-goat Ig peroxidase kit) was applied to the blood smears for 30 min. Lastly, Vector NovaRED, which is a substrate for peroxidase, was applied for 5 min (Vector Laboratories; #SK-4800), and a color change in red served as the reporter for antibody binding to PSGL-1.

    Techniques: Labeling, Plasmid Preparation