Histology was performed by HistoWiz Inc. (histowiz.com ) using a Standard Operating Procedure and fully automated workflow. Samples were processed, embedded in paraffin, and sectioned at 4 μm. Immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1000) using standard protocols. Antibodies used were rat monoclonal F4/80 primary antibody (eBioscience; 14‐4801; 1:200) and rabbit anti‐rat secondary (Vector; 1:100). Bond Polymer Refine Detection (Leica Biosystems) was used according to the manufacturer's protocol. After staining, sections were dehydrated and film coverslipped using a TissueTek‐Prisma and Coverslipper (Sakura). Whole slide scanning (x40) was performed on an Aperio AT2 (Leica Biosystems). Four separate lots of dehydrated amniotic membrane and four separate lots of dehydrated amnion/chorion were sent in biopsy cages with 50% paraformaldehyde/PBS. Stains include H&E for structure and nuclei, Van Gieson for elastin and collagen, Alcian blue for polysaccharides, and immunohistochemistry for collagen I, collagen III, fibronectin, and laminin.
Rabbit anti rat secondary
Manufactured by Vector Laboratories
Rabbit anti-rat secondary is a laboratory reagent used in immunoassays and other immunological techniques. It is a purified antibody that specifically binds to rat primary antibodies, allowing their detection and quantification.
Automatically generated - may contain errors
Lab products found in correlation
Aperio at2, by Leica (8 mentions)
Bond polymer refine detection, by Leica (8 mentions)
Bond rx, by Leica (6 mentions)
F4 80 primary antibody, by Thermo Fisher Scientific (3 mentions)
Rat monoclonal f4 80 primary antibody, by Thermo Fisher Scientific (3 mentions)
Bond rx autostainer, by Leica (1 mentions)
Ht501128, by Merck Group (1 mentions)
8 protocols using rabbit anti rat secondary
1
Histological Characterization of Amniotic Membranes
2
Histopathological Analysis of Mice Tissues
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Mice were euthanized and tissue samples (tumor, liver and kidney) were excised from mice immediately, washed with PBS and placed in 10% buffered formalin for 24 h and transferred to 70% ethanol for histopathological analysis. Histology was performed by HistoWiz Inc. (histowiz.com ) using a Standard Operating Procedure and fully automated workflow. Samples were processed, embedded in paraffin, and sectioned at 4 μm (Harder et al., 2009 (link); Pignochino et al., 2010 (link); Walsh et al., 2013 (link)). Immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1000) using standard protocols. Antibodies used were rat monoclonal F4/80 primary antibody (eBioscience, 14–4801, and 1:200) and rabbit anti-rat secondary (Vector, 1:100) (Harder et al., 2009 (link); Pignochino et al., 2010 (link); Walsh et al., 2013 (link)). Bond Polymer Refine Detection (Leica Biosystems) was used according to manufacturer's protocol. After staining, sections were dehydrated and film coverslipped using a Tissue-Tek Prisma and Coverslipper (Sakura). Whole slide scanning (40×) was performed on an Aperio AT2 (Leica Biosystems).
3
Histopathological Analysis of Tumor Samples
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Tissue samples from mice bearing tumors from Black and White PCa patients post treatments with GemHCl and 4NSG-SLN were excised and immediately washed with PBS, fixed (10% buffered formalin) for 24 h, and transferred to 70% ethanol for histopathological analysis. Histology was performed by HistoWiz Inc. (Histowiz.com) according to manufacturer protocol. Samples were embedded in paraffin, and 4 μm sections were prepared [7 (link), 40 (link), 41 (link)]. Immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1000) using standard protocols (https://home.histowiz.com/faq/ ). Samples were incubated overnight with primary antibodies (Antibodies used were rat monoclonal F4/80 primary antibody (eBioscience, 14–4801, and 1:200), and immunohistochemical staining was done using rabbit anti-rat secondary (Vector Labs, 1:100) [7 (link), 40 (link), 41 (link)]. Bond Polymer Refine Detection was used per the manufacturer's protocol (Leica Biosystems). After staining, sections were dehydrated and filmed, coverslipped using a Tissue-Tek Prisma coverslip (Sakura). Whole slide scanning (40x) was performed on an Aperio AT2 (Leica Biosystems).
4
Immunohistochemical Analysis of Tumor Samples
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Tumor samples excised from the mice, washed with PBS, placed in 10% buffered formalin for 24 h, and transferred to 70% ethanol for histopathological analysis. Immunohistochemistry was performed by Histowiz Inc. (histowiz.com ) using a Standard Operating Procedure and fully automated workflow. Samples were processed, embedded in paraffin, and sectioned at 4μm [16 (link)]. Immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1000) using standard protocols. Antibodies used were rat monoclonal F4/80 primary antibody (eBioscience, 14–4801, and 1:200) and rabbit anti-rat secondary (Vector, 1:100) [17 (link)–19 (link)]. Bond Polymer Refine Detection (Leica Biosystems) was used according to the manufacturer’s protocol. After staining, sections were dehydrated, and film cover slipped using a Tissue-Tek Prisma and Cover-slipper (Sakura). Whole slide scanning (40×) was performed on an Aperio AT2 (Leica Biosystems).
5
Histopathological Analysis of Tumor, Liver, and Kidney Tissues
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Mice were euthanized and tissue samples (tumor, liver and kidney) were excised from mice immediately, washed with PBS and placed in 10% buffered formalin for 24 h and transferred to 70% ethanol for histopathological analysis. Histology was performed by HistoWiz Inc. (histowiz.com) using a Standard Operating Procedure and fully automated workflow. Samples were processed, embedded in paraffin, and sectioned at 4 μm52 (link)–54 (link). Immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1,000) using standard protocols. Antibodies used were rat monoclonal F4/80 primary antibody (eBioscience, 14-4801, and 1:200) and rabbit anti-rat secondary (Vector, 1:100)52 (link)–54 (link). Bond Polymer Refine Detection (Leica Biosystems) was used according to manufacturer’s protocol. After staining, sections were dehydrated and film coverslipped using a Tissue-Tek Prisma and Coverslipper (Sakura). Whole slide scanning (40 ×) was performed on an Aperio AT2 (Leica Biosystems).
6
Automated Immunohistochemistry Workflow for Quantitative Analysis
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Immunohistochemistry was performed by HistoWiz Inc. (histowiz.com ) using a Standard Operating Procedure and fully automated workflow. Samples were processed, embedded in paraffin, and sectioned at 4μm. Immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1000) using standard protocols. Antibodies used were rat monoclonal F4/80 primary antibody (eBioscience, 14–4801, 1:200) and rabbit anti-rat secondary (Vector, 1:100). Bond Polymer Refine Detection (Leica Biosystems) was used according to the manufacturer’s protocol. After staining, sections were dehydrated and film coverslipped using a TissueTek-Prisma and Coverslipper (Sakura). Whole slide scanning (40x) was performed on an Aperio AT2 (Leica Biosystems). Qupath was used to measure overall DAB staining, annotate the area of the mammary gland, and to perform cell segmentation. For each cell, Qupath measures a variety of morphology parameters including nuclear area, perimeter, and max calipers. Downstream analysis of the segmented cells was done in Python.
7
Skin Immunohistochemistry Protocol for Neonates
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Back skin dissected from neonates was fixed in PBS with 4% PFA at 4°C overnight. Then the tissue was washed in PBS with 0%, 35% and 70% EtOH gradually and sent to Histowiz for further processing. Generally, immunohistochemistry was performed on a Bond Rx autostainer (Leica Biosystems) with enzyme treatment (1:1000) using standard protocols. Antibodies used were rat monoclonal F4/80 primary antibody (1:200, eBioscience, 14–4801) and rabbit anti-rat secondary (1:100, Vector). Bond Polymer Refine Detection (Leica Biosystems) was used according to manufacturer’s protocol. After staining, sections were dehydrated and film coverslipped using a TissueTek-Prisma and Coverslipper (Sakura). Whole slide scanning (40x) was performed on an Aperio AT2 (Leica Biosystems).
8
Immunohistochemical Tumor Tissue Analysis
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