immunoperoxidase kit  (Vector Laboratories)


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    Name:
    VECTASTAIN ABC AP Staining KIT Alkaline Phosphatase Standard
    Description:
    VECTASTAIN ABC AP Staining Kits remains widely popular with more than 50 000 citations to its credit Based on the versatile avidin biotin complex interaction the system is modular and along with our selection of secondary antibodies can accommodate a wide array of primary antibody and tissue species Our ABC kits are economical and continue to be a staple product in any immunohistochemistry IHC and immunocytochemistry ICC laboratory The sensitivity of the VECTASTAIN ABC AP system is comparable to that of the peroxidase VECTASTAIN Elite ABC system The VECTASTAIN ABC AP Kits may be preferred for tissues that have high endogenous peroxidase activity The system also offers additional substrate color choices VECTASTAIN ABC AP Kit Components 2 ml Reagent A2 ml Reagent BReagent A and B when mixed form the ABC AP complex Kit is sufficient to stain approximately 1000 2000 sections or fifty 100 cm2 blots
    Catalog Number:
    AK-5000
    Price:
    None
    Category:
    Protein chromogenic detection reagents or kits or substrates
    Reactivity:
    No antibody included
    Size:
    1 Kit
    Buy from Supplier


    Structured Review

    Vector Laboratories immunoperoxidase kit
    VECTASTAIN ABC AP Staining KIT Alkaline Phosphatase Standard
    VECTASTAIN ABC AP Staining Kits remains widely popular with more than 50 000 citations to its credit Based on the versatile avidin biotin complex interaction the system is modular and along with our selection of secondary antibodies can accommodate a wide array of primary antibody and tissue species Our ABC kits are economical and continue to be a staple product in any immunohistochemistry IHC and immunocytochemistry ICC laboratory The sensitivity of the VECTASTAIN ABC AP system is comparable to that of the peroxidase VECTASTAIN Elite ABC system The VECTASTAIN ABC AP Kits may be preferred for tissues that have high endogenous peroxidase activity The system also offers additional substrate color choices VECTASTAIN ABC AP Kit Components 2 ml Reagent A2 ml Reagent BReagent A and B when mixed form the ABC AP complex Kit is sufficient to stain approximately 1000 2000 sections or fifty 100 cm2 blots
    https://www.bioz.com/result/immunoperoxidase kit/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoperoxidase kit - by Bioz Stars, 2021-06
    99/100 stars

    Images

    1) Product Images from "Analysis of Pathogen-Host Cell Interactions in Purpura Fulminans: Expression of Capsule, Type IV Pili, and PorA by Neisseria meningitidis In Vivo "

    Article Title: Analysis of Pathogen-Host Cell Interactions in Purpura Fulminans: Expression of Capsule, Type IV Pili, and PorA by Neisseria meningitidis In Vivo

    Journal: Infection and Immunity

    doi: 10.1128/IAI.70.9.5193-5201.2002

    Immunohistochemical staining of inflammatory cells and the vascular endothelium in skin biopsy samples from patients with meningococcal disease. The inflammatory infiltrate consisted of a mixture of CD68-positive macrophages (A) (magnification, ×400) and neutrophil elastase-positive polymorphonuclear cells (B) (magnification, ×400). Multiple CD31-positive blood vessels were seen throughout the biopsy samples (C and D) (magnification, ×400 and 600, respectively). Arrows indicate positive immunoperoxidase staining (brown) with the appropriate specific mouse monoclonal antibody (nuclei were counterstained with hematoxylin).
    Figure Legend Snippet: Immunohistochemical staining of inflammatory cells and the vascular endothelium in skin biopsy samples from patients with meningococcal disease. The inflammatory infiltrate consisted of a mixture of CD68-positive macrophages (A) (magnification, ×400) and neutrophil elastase-positive polymorphonuclear cells (B) (magnification, ×400). Multiple CD31-positive blood vessels were seen throughout the biopsy samples (C and D) (magnification, ×400 and 600, respectively). Arrows indicate positive immunoperoxidase staining (brown) with the appropriate specific mouse monoclonal antibody (nuclei were counterstained with hematoxylin).

    Techniques Used: Immunohistochemistry, Staining, Immunoperoxidase Staining

    Related Articles

    Staining:

    Article Title: Immunohistochemical analysis of macroautophagy
    Article Snippet: In case the primary antibody was biotinylated, blocking with normal serum and incubation with biotinylated secondary antibodies were omitted. .. In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase. .. The procedure is similar with the staining steps for frozen sections described above, except quenching of endogenous peroxidase is no longer required.

    Article Title: Induction of cyclooxygenase-2 in monocyte/macrophage by mucins secreted from colon cancer cells
    Article Snippet: .. Immunohistochemical staining was performed with the Vectastain avidin–biotin peroxidase complex kit (Vector Laboratories) as described ( ). .. Double immunostaining was carried out according to Kawahito et al. ( ).

    Article Title: Diet-induced obesity links to ER positive breast cancer progression via LPA/PKD-1-CD36 signaling-mediated microvascular remodeling
    Article Snippet: E0771 cells (1×106 cells/mouse) were implanted into mice close to the fourth mammary pad [ ] and tumor volume was measured using calipers in two dimensions and calculated using the formula: (width2 × length)/2 [ ]. .. Immunofluorescence and immunohistochemical assays Tissue immunohistochemical and immunofluorescence staining was performed using antibodies and methods as described in our previous studies [ , , ] or by using the Vectastain® ABC kit (Vector Laboratories). .. Mouse angiogenesis profiling A Proteome Profiler™ antibody array (R & D system® ) was performed to determine relative protein levels of angiogenic factors in the plasma according to the manufacturer's instruction.

    Article Title: Dynamic Populations of Dendritic Cell-Specific ICAM-3 Grabbing Nonintegrin-Positive Immature Dendritic Cells and Liver/Lymph Node-Specific ICAM-3 Grabbing Nonintegrin-Positive Endothelial Cells in the Outer Zones of the Paracortex of Human Lymph Nodes
    Article Snippet: Paraffin sections were rehydrated and subjected to antigen-retrieval by boiling in 0.01 mol/L citric acid (pH 6.0) for 10 minutes before incubation with antibodies. .. Staining was performed with the ABC-AP Vectastain kit (Vector Laboratories) or ABC-PO and diaminobenzidine tetrahydrochloride (0.5 mg/ml) and sections were counterstained with hematoxylin according to Pappanicolau. .. For immunofluorescence, Alexa 488 (Molecular Probes, Eugene, OR)- and Texas Red (Jackson, West Grove, PA)-conjugated secondary antibodies were used.

    Labeling:

    Article Title: Analysis of Pathogen-Host Cell Interactions in Purpura Fulminans: Expression of Capsule, Type IV Pili, and PorA by Neisseria meningitidis In Vivo
    Article Snippet: Briefly, sections were incubated in 0.3% aqueous hydrogen peroxide to quench endogenous peroxidase activity and blocked with normal mouse or rabbit serum before incubation with the monoclonal antibodies for 1 h at room temperature. .. Labeling was detected with an immunoperoxidase kit (Vectastain Elite ABC kit; Vector Laboratories, Burlingame, Calif.), and sections were counterstained in hematoxylin. .. Antigen retrieval treatment of the sections was required for optimal staining with all of the meningococcal monoclonal antibodies and the anti-CD31 and -CD68 antibodies ( ).

    Incubation:

    Article Title: Luteolin Inhibits Human Prostate Tumor Growth by Suppressing Vascular Endothelial Growth Factor Receptor 2-Mediated Angiogenesis
    Article Snippet: Histology and Immunohistochemistry Tumor tissues were fixed in 10% neutral-buffered formalin for 24 hours, processed, and embedded in paraffin blocks. .. The sections (5 µm) were blocked with 10% goat serum and incubated with a rabbit anti-CD31 (1∶100; Novus Biologicals Inc, Littleton, CO) and mouse ant-CD34 (1∶100; BD Pharmingen Inc, San Diego, CA) antibodies for 24 h. The slides were subsequently incubated for 30 min with biotinylated anti-rabbit/anti-mouse secondary antibody (Vector laboratories, Burlingame, CA) and followed by incubation of Vectastain ABC Kit (Vector Laboratories). .. Diaminobenzidine (Sigma) was used as the chromagen and methyl green (Sigma) as the counterstain.

    Immunohistochemistry:

    Article Title: Induction of cyclooxygenase-2 in monocyte/macrophage by mucins secreted from colon cancer cells
    Article Snippet: .. Immunohistochemical staining was performed with the Vectastain avidin–biotin peroxidase complex kit (Vector Laboratories) as described ( ). .. Double immunostaining was carried out according to Kawahito et al. ( ).

    Article Title: Diet-induced obesity links to ER positive breast cancer progression via LPA/PKD-1-CD36 signaling-mediated microvascular remodeling
    Article Snippet: E0771 cells (1×106 cells/mouse) were implanted into mice close to the fourth mammary pad [ ] and tumor volume was measured using calipers in two dimensions and calculated using the formula: (width2 × length)/2 [ ]. .. Immunofluorescence and immunohistochemical assays Tissue immunohistochemical and immunofluorescence staining was performed using antibodies and methods as described in our previous studies [ , , ] or by using the Vectastain® ABC kit (Vector Laboratories). .. Mouse angiogenesis profiling A Proteome Profiler™ antibody array (R & D system® ) was performed to determine relative protein levels of angiogenic factors in the plasma according to the manufacturer's instruction.

    Avidin-Biotin Assay:

    Article Title: Induction of cyclooxygenase-2 in monocyte/macrophage by mucins secreted from colon cancer cells
    Article Snippet: .. Immunohistochemical staining was performed with the Vectastain avidin–biotin peroxidase complex kit (Vector Laboratories) as described ( ). .. Double immunostaining was carried out according to Kawahito et al. ( ).

    Article Title: Bone Matrix Proteins: Isolation and Characterization of a Novel Cell-binding Keratan Sulfate Proteoglycan (Osteoadherin) from Bovine Bone
    Article Snippet: The sections were incubated with primary antibody against osteoadherin (diluted to 1:1,000 in PBS-0.01% BSA) or the preimmune serum (diluted to 1:1,000 in PBS 0.01% BSA) at 4°C overnight in a moist chamber. .. The sections were then treated with biotinylated secondary antibody (diluted 1:200) and avidin–peroxidase conjugate using the Vectastain ABC kit™ (Vector Labs, Burlingame, CA), following the recommendations of the manufacturer. .. Cell Attachment to Osteoadherin Primary osteoblasts isolated with the method of Robey et al. ( ) were grown to near confluency in Ham's F12 medium supplemented with 10% FBS, 50 UI penicillin, and 50 μg/ml streptomycin.

    Immunofluorescence:

    Article Title: Diet-induced obesity links to ER positive breast cancer progression via LPA/PKD-1-CD36 signaling-mediated microvascular remodeling
    Article Snippet: E0771 cells (1×106 cells/mouse) were implanted into mice close to the fourth mammary pad [ ] and tumor volume was measured using calipers in two dimensions and calculated using the formula: (width2 × length)/2 [ ]. .. Immunofluorescence and immunohistochemical assays Tissue immunohistochemical and immunofluorescence staining was performed using antibodies and methods as described in our previous studies [ , , ] or by using the Vectastain® ABC kit (Vector Laboratories). .. Mouse angiogenesis profiling A Proteome Profiler™ antibody array (R & D system® ) was performed to determine relative protein levels of angiogenic factors in the plasma according to the manufacturer's instruction.

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  • 99
    Vector Laboratories immunoperoxidase kit
    Immunohistochemical staining of inflammatory cells and the vascular endothelium in skin biopsy samples from patients with meningococcal disease. The inflammatory infiltrate consisted of a mixture of CD68-positive macrophages (A) (magnification, ×400) and neutrophil elastase-positive polymorphonuclear cells (B) (magnification, ×400). Multiple CD31-positive blood vessels were seen throughout the biopsy samples (C and D) (magnification, ×400 and 600, respectively). Arrows indicate positive <t>immunoperoxidase</t> staining (brown) with the appropriate specific mouse monoclonal antibody (nuclei were counterstained with hematoxylin).
    Immunoperoxidase Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoperoxidase kit/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoperoxidase kit - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    86
    Vector Laboratories mouse rat specific immunoperoxidase kit
    Effect of H-7 and H-8 on IL-1α induced migration of LC. Following hair removal and death, dorsal skin sections were harvested from paired normal BALB/c mice. Sections were placed (epidermis up) on medium moistened filters prior to addition of either 25 μl of H-7 (100 μg/ml in DMSO, H-8 (100 μg/ml in DMSO) or DMSO alone. Two hours later, designated sections received either IL-1α (5 ng/ml in DMSO) or DMSO alone, after which skin sections were cultured at 37° in the presence of 5% CO 2 . After 48 hr of culture, epidermal sheets were removed from each section, stained for IA by <t>immunoperoxidase</t> and the numbers of LC counted with the aid of a microscope.
    Mouse Rat Specific Immunoperoxidase Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse rat specific immunoperoxidase kit/product/Vector Laboratories
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse rat specific immunoperoxidase kit - by Bioz Stars, 2021-06
    86/100 stars
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    96
    Vector Laboratories peroxidase substrates aminoethylcarbazole
    Immunohistochemical demonstration of ERK/MAPK phosphorylation in astrocytic neoplasms. For each tumor, adjacent sections were immunolabeled with the phospho-specific ERK/MAPK antibody (pMAPK; left panels ) and a phospho-insensitive ERK/MAPK antibody (MAPK; right panels ). Elevated nuclear and cytoplasmic dp-ERK/MAPK immunoreactivity, relative to normal quiescent astrocytes, was detected in a subset of tumor cells within a variety of astrocytic neoplasms. In contrast, ERK/MAPK immunoreactivity was more uniformly expressed. A and B: Astrocytoma, WHO grade II. Some diffusely infiltrating astrocytoma cells show increased nuclear and cytoplasmic dp-ERK/MAPK immunoreactivity. Note the absence of labeling in adjacent normal neurons. ERK/MAPK protein is more evenly expressed in tumor cell cytoplasm and in neurons. C and D: Anaplastic astrocytoma, WHO grade III. A subset of gemistocytic tumor cells shows high cytoplasmic and nuclear dp-ERK/MAPK immunoreactivity. E and F: Ependymoma. A subpopulation of highly fibrillar ependymoma cells, especially those forming perivascular pseudorosettes, shows elevated dp-ERK/MAPK immunoreactivity; compare with the relatively uniform expression of ERK/MAPK protein. G and H: Glioblastoma multiforme (astrocytoma, WHO grade IV). Tumor cells near the hyperplastic microvasculature show the most intense nuclear and cytoplasmic pMAPK immunoreactivity. Some fine immunoreactive tumor cell processes appear to be oriented toward the microvasculature ( corner inset ). Total MAPK immunoreactivity is much more uniform, and not restricted to the perivascular tumor cells. I and J: Glioblastoma with giant cell transformation (astrocytoma, WHO grade IV). Only the large, bizarre, multinucleated tumor cells have detectable dp-ERK/MAPK immunoreactivity; the small cell population is completely negative. Note the absence of dp-ERK/MAPK immunoreactivity in a mitotic tumor cell ( left side ). In contrast, both small and large tumor cell populations show uniform ERK/MAPK protein immunoreactivity. Also, a mitotic cell is visible that has detectable MAPK immunoreactivity ( right side ). K to N: Selective dephosphorylation of ERK/MAPK within mitotic and cycling glioblastoma cells. K and L: Immunoreactivity for dp-ERK/MAPK is absent within individual mitotic tumor cells in two additional glioblastoma specimens; adjacent interphase cells show abundant cytoplasmic and nuclear immunoreactivity. M: Evidence that mitotic phosphoepitopes in general are not lost in routinely processed surgical specimens comes from immunolabeling with monoclonal antibody MPM-2, which recognizes a number of mitosis-specific phosphorylated proteins. Strong perichromosomal staining is present in mitotic cells. N: Double-label immunohistochemistry for dp-ERK/MAPK (red-brown) and Ki-67 (dark blue) reveals essentially complete non-co-localization. This suggests that cycling cells (those with nuclear Ki-67 immunolabeling) and those with activated ERK/MAPK (dp-ERK/MAPK immunoreactive) are generally mutually exclusive populations. A to M: Immunoperoxidase staining (DAB) with hematoxylin counterstain. N: Double-label immunoperoxidase with <t>aminoethylcarbazole</t> and Vector SG substrates; no counterstain. Scale bar in A represents 50 μm and applies to A to J and M ; bar in N represents 20 μm and applies to K , L , and N .
    Peroxidase Substrates Aminoethylcarbazole, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peroxidase substrates aminoethylcarbazole/product/Vector Laboratories
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peroxidase substrates aminoethylcarbazole - by Bioz Stars, 2021-06
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    94
    Vector Laboratories avidin biotin immunoperoxidase reaction
    Immunohistochemical localization of AT2 and AT1 in the medial layer of the aorta. The immunostaining, using antibodies for von Willebrand factor (vWF) ( a ) and VSM-specific α-actin ( b ), indicates the localization of the endothelium and medial layer in the aorta, respectively. Positive immunostaining for AT2 was highly localized in the medial layer of the aorta from AT2-TG mice ( c ), whereas in the wild-type mice, no significant AT2 signals were observed in the aorta ( d ). Immunochemical signals for AT1 were detected in the medial layers of the aortas from wild-type and AT2-TG mice to a similar extent ( e and f ). Positive staining was viewed with avidin-biotin <t>immunoperoxidase</t> reaction using diaminobenzidine. Asterisks indicate the lumen of the aorta.
    Avidin Biotin Immunoperoxidase Reaction, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/avidin biotin immunoperoxidase reaction/product/Vector Laboratories
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    avidin biotin immunoperoxidase reaction - by Bioz Stars, 2021-06
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    Image Search Results


    Immunohistochemical staining of inflammatory cells and the vascular endothelium in skin biopsy samples from patients with meningococcal disease. The inflammatory infiltrate consisted of a mixture of CD68-positive macrophages (A) (magnification, ×400) and neutrophil elastase-positive polymorphonuclear cells (B) (magnification, ×400). Multiple CD31-positive blood vessels were seen throughout the biopsy samples (C and D) (magnification, ×400 and 600, respectively). Arrows indicate positive immunoperoxidase staining (brown) with the appropriate specific mouse monoclonal antibody (nuclei were counterstained with hematoxylin).

    Journal: Infection and Immunity

    Article Title: Analysis of Pathogen-Host Cell Interactions in Purpura Fulminans: Expression of Capsule, Type IV Pili, and PorA by Neisseria meningitidis In Vivo

    doi: 10.1128/IAI.70.9.5193-5201.2002

    Figure Lengend Snippet: Immunohistochemical staining of inflammatory cells and the vascular endothelium in skin biopsy samples from patients with meningococcal disease. The inflammatory infiltrate consisted of a mixture of CD68-positive macrophages (A) (magnification, ×400) and neutrophil elastase-positive polymorphonuclear cells (B) (magnification, ×400). Multiple CD31-positive blood vessels were seen throughout the biopsy samples (C and D) (magnification, ×400 and 600, respectively). Arrows indicate positive immunoperoxidase staining (brown) with the appropriate specific mouse monoclonal antibody (nuclei were counterstained with hematoxylin).

    Article Snippet: Labeling was detected with an immunoperoxidase kit (Vectastain Elite ABC kit; Vector Laboratories, Burlingame, Calif.), and sections were counterstained in hematoxylin.

    Techniques: Immunohistochemistry, Staining, Immunoperoxidase Staining

    Effect of H-7 and H-8 on IL-1α induced migration of LC. Following hair removal and death, dorsal skin sections were harvested from paired normal BALB/c mice. Sections were placed (epidermis up) on medium moistened filters prior to addition of either 25 μl of H-7 (100 μg/ml in DMSO, H-8 (100 μg/ml in DMSO) or DMSO alone. Two hours later, designated sections received either IL-1α (5 ng/ml in DMSO) or DMSO alone, after which skin sections were cultured at 37° in the presence of 5% CO 2 . After 48 hr of culture, epidermal sheets were removed from each section, stained for IA by immunoperoxidase and the numbers of LC counted with the aid of a microscope.

    Journal: Immunology

    Article Title: Protein-kinase-specific inhibitors block Langerhans' cell migration by inhibiting interleukin-1? release

    doi: 10.1046/j.1365-2567.1999.00680.x

    Figure Lengend Snippet: Effect of H-7 and H-8 on IL-1α induced migration of LC. Following hair removal and death, dorsal skin sections were harvested from paired normal BALB/c mice. Sections were placed (epidermis up) on medium moistened filters prior to addition of either 25 μl of H-7 (100 μg/ml in DMSO, H-8 (100 μg/ml in DMSO) or DMSO alone. Two hours later, designated sections received either IL-1α (5 ng/ml in DMSO) or DMSO alone, after which skin sections were cultured at 37° in the presence of 5% CO 2 . After 48 hr of culture, epidermal sheets were removed from each section, stained for IA by immunoperoxidase and the numbers of LC counted with the aid of a microscope.

    Article Snippet: LC were then enumerated by immunoperoxidase staining using a mouse/ rat specific immunoperoxidase kit (Vector Laboratories, Burlingame, CA).

    Techniques: Migration, Mouse Assay, Cell Culture, Staining, IA, Microscopy

    Immunohistochemical demonstration of ERK/MAPK phosphorylation in astrocytic neoplasms. For each tumor, adjacent sections were immunolabeled with the phospho-specific ERK/MAPK antibody (pMAPK; left panels ) and a phospho-insensitive ERK/MAPK antibody (MAPK; right panels ). Elevated nuclear and cytoplasmic dp-ERK/MAPK immunoreactivity, relative to normal quiescent astrocytes, was detected in a subset of tumor cells within a variety of astrocytic neoplasms. In contrast, ERK/MAPK immunoreactivity was more uniformly expressed. A and B: Astrocytoma, WHO grade II. Some diffusely infiltrating astrocytoma cells show increased nuclear and cytoplasmic dp-ERK/MAPK immunoreactivity. Note the absence of labeling in adjacent normal neurons. ERK/MAPK protein is more evenly expressed in tumor cell cytoplasm and in neurons. C and D: Anaplastic astrocytoma, WHO grade III. A subset of gemistocytic tumor cells shows high cytoplasmic and nuclear dp-ERK/MAPK immunoreactivity. E and F: Ependymoma. A subpopulation of highly fibrillar ependymoma cells, especially those forming perivascular pseudorosettes, shows elevated dp-ERK/MAPK immunoreactivity; compare with the relatively uniform expression of ERK/MAPK protein. G and H: Glioblastoma multiforme (astrocytoma, WHO grade IV). Tumor cells near the hyperplastic microvasculature show the most intense nuclear and cytoplasmic pMAPK immunoreactivity. Some fine immunoreactive tumor cell processes appear to be oriented toward the microvasculature ( corner inset ). Total MAPK immunoreactivity is much more uniform, and not restricted to the perivascular tumor cells. I and J: Glioblastoma with giant cell transformation (astrocytoma, WHO grade IV). Only the large, bizarre, multinucleated tumor cells have detectable dp-ERK/MAPK immunoreactivity; the small cell population is completely negative. Note the absence of dp-ERK/MAPK immunoreactivity in a mitotic tumor cell ( left side ). In contrast, both small and large tumor cell populations show uniform ERK/MAPK protein immunoreactivity. Also, a mitotic cell is visible that has detectable MAPK immunoreactivity ( right side ). K to N: Selective dephosphorylation of ERK/MAPK within mitotic and cycling glioblastoma cells. K and L: Immunoreactivity for dp-ERK/MAPK is absent within individual mitotic tumor cells in two additional glioblastoma specimens; adjacent interphase cells show abundant cytoplasmic and nuclear immunoreactivity. M: Evidence that mitotic phosphoepitopes in general are not lost in routinely processed surgical specimens comes from immunolabeling with monoclonal antibody MPM-2, which recognizes a number of mitosis-specific phosphorylated proteins. Strong perichromosomal staining is present in mitotic cells. N: Double-label immunohistochemistry for dp-ERK/MAPK (red-brown) and Ki-67 (dark blue) reveals essentially complete non-co-localization. This suggests that cycling cells (those with nuclear Ki-67 immunolabeling) and those with activated ERK/MAPK (dp-ERK/MAPK immunoreactive) are generally mutually exclusive populations. A to M: Immunoperoxidase staining (DAB) with hematoxylin counterstain. N: Double-label immunoperoxidase with aminoethylcarbazole and Vector SG substrates; no counterstain. Scale bar in A represents 50 μm and applies to A to J and M ; bar in N represents 20 μm and applies to K , L , and N .

    Journal: The American Journal of Pathology

    Article Title: In Situ Visualization of Intratumor Growth Factor Signaling

    doi:

    Figure Lengend Snippet: Immunohistochemical demonstration of ERK/MAPK phosphorylation in astrocytic neoplasms. For each tumor, adjacent sections were immunolabeled with the phospho-specific ERK/MAPK antibody (pMAPK; left panels ) and a phospho-insensitive ERK/MAPK antibody (MAPK; right panels ). Elevated nuclear and cytoplasmic dp-ERK/MAPK immunoreactivity, relative to normal quiescent astrocytes, was detected in a subset of tumor cells within a variety of astrocytic neoplasms. In contrast, ERK/MAPK immunoreactivity was more uniformly expressed. A and B: Astrocytoma, WHO grade II. Some diffusely infiltrating astrocytoma cells show increased nuclear and cytoplasmic dp-ERK/MAPK immunoreactivity. Note the absence of labeling in adjacent normal neurons. ERK/MAPK protein is more evenly expressed in tumor cell cytoplasm and in neurons. C and D: Anaplastic astrocytoma, WHO grade III. A subset of gemistocytic tumor cells shows high cytoplasmic and nuclear dp-ERK/MAPK immunoreactivity. E and F: Ependymoma. A subpopulation of highly fibrillar ependymoma cells, especially those forming perivascular pseudorosettes, shows elevated dp-ERK/MAPK immunoreactivity; compare with the relatively uniform expression of ERK/MAPK protein. G and H: Glioblastoma multiforme (astrocytoma, WHO grade IV). Tumor cells near the hyperplastic microvasculature show the most intense nuclear and cytoplasmic pMAPK immunoreactivity. Some fine immunoreactive tumor cell processes appear to be oriented toward the microvasculature ( corner inset ). Total MAPK immunoreactivity is much more uniform, and not restricted to the perivascular tumor cells. I and J: Glioblastoma with giant cell transformation (astrocytoma, WHO grade IV). Only the large, bizarre, multinucleated tumor cells have detectable dp-ERK/MAPK immunoreactivity; the small cell population is completely negative. Note the absence of dp-ERK/MAPK immunoreactivity in a mitotic tumor cell ( left side ). In contrast, both small and large tumor cell populations show uniform ERK/MAPK protein immunoreactivity. Also, a mitotic cell is visible that has detectable MAPK immunoreactivity ( right side ). K to N: Selective dephosphorylation of ERK/MAPK within mitotic and cycling glioblastoma cells. K and L: Immunoreactivity for dp-ERK/MAPK is absent within individual mitotic tumor cells in two additional glioblastoma specimens; adjacent interphase cells show abundant cytoplasmic and nuclear immunoreactivity. M: Evidence that mitotic phosphoepitopes in general are not lost in routinely processed surgical specimens comes from immunolabeling with monoclonal antibody MPM-2, which recognizes a number of mitosis-specific phosphorylated proteins. Strong perichromosomal staining is present in mitotic cells. N: Double-label immunohistochemistry for dp-ERK/MAPK (red-brown) and Ki-67 (dark blue) reveals essentially complete non-co-localization. This suggests that cycling cells (those with nuclear Ki-67 immunolabeling) and those with activated ERK/MAPK (dp-ERK/MAPK immunoreactive) are generally mutually exclusive populations. A to M: Immunoperoxidase staining (DAB) with hematoxylin counterstain. N: Double-label immunoperoxidase with aminoethylcarbazole and Vector SG substrates; no counterstain. Scale bar in A represents 50 μm and applies to A to J and M ; bar in N represents 20 μm and applies to K , L , and N .

    Article Snippet: Double-label immunohistochemistry was performed sequentially using the peroxidase substrates aminoethylcarbazole and Vector SG, according to the manufacturer’s instructions (Vector Laboratories, Burlingame, CA).

    Techniques: Immunohistochemistry, Immunolabeling, Labeling, Expressing, Transformation Assay, De-Phosphorylation Assay, Staining, Immunoperoxidase Staining, Plasmid Preparation

    Immunohistochemical localization of AT2 and AT1 in the medial layer of the aorta. The immunostaining, using antibodies for von Willebrand factor (vWF) ( a ) and VSM-specific α-actin ( b ), indicates the localization of the endothelium and medial layer in the aorta, respectively. Positive immunostaining for AT2 was highly localized in the medial layer of the aorta from AT2-TG mice ( c ), whereas in the wild-type mice, no significant AT2 signals were observed in the aorta ( d ). Immunochemical signals for AT1 were detected in the medial layers of the aortas from wild-type and AT2-TG mice to a similar extent ( e and f ). Positive staining was viewed with avidin-biotin immunoperoxidase reaction using diaminobenzidine. Asterisks indicate the lumen of the aorta.

    Journal: Journal of Clinical Investigation

    Article Title: Angiotensin II type 2 receptor overexpression activates the vascular kinin system and causes vasodilation

    doi:

    Figure Lengend Snippet: Immunohistochemical localization of AT2 and AT1 in the medial layer of the aorta. The immunostaining, using antibodies for von Willebrand factor (vWF) ( a ) and VSM-specific α-actin ( b ), indicates the localization of the endothelium and medial layer in the aorta, respectively. Positive immunostaining for AT2 was highly localized in the medial layer of the aorta from AT2-TG mice ( c ), whereas in the wild-type mice, no significant AT2 signals were observed in the aorta ( d ). Immunochemical signals for AT1 were detected in the medial layers of the aortas from wild-type and AT2-TG mice to a similar extent ( e and f ). Positive staining was viewed with avidin-biotin immunoperoxidase reaction using diaminobenzidine. Asterisks indicate the lumen of the aorta.

    Article Snippet: Staining was viewed with the avidin-biotin immunoperoxidase reaction (VECTASTAIN ABC Kit; Vector Laboratories, Burlingame, California, USA) using diaminobenzidine (Fast DAB tablets; Sigma).

    Techniques: Immunohistochemistry, Immunostaining, Mouse Assay, Staining, Avidin-Biotin Assay