CD163 has been used as a specific marker to identify M2 macrophages [13 , 14 (link)]. Surgically resected specimens were retrieved to perform immunohistochemistry. Sections 4 μm in thickness were deparaffined and rehydrated. The sections were then subjected to endogenous peroxidase blocking in 1% H2O2 solution in methanol for 15 min. Antigen retrieval was performed by autoclaving the sections at 105 °C for 10 min in Dako Target Retrieval Solution (Dako, Glostrup, Denmark). Serum blocking was performed with 10% normal rabbit serum for 10 min. After H2O2 and serum blocking, the slides were incubated with primary mouse monoclonal anti-CD163 antibody (1:200 dilution; Leica Biosystems, Newcastle Upon Tyne, UK) at room temperature for 1 h. The secondary antibody was biotin-labeled rabbit anti-mouse IgG (1:500; Nichirei, Tokyo, Japan). Detection was performed with a DAB kit (Histofine simple stain kit; Nichirei). The sections were counterstained with hematoxylin.
Biotin labeled rabbit anti mouse igg
Manufactured by Nichirei Biosciences
Sourced in Japan
Biotin-labeled rabbit anti-mouse IgG is a secondary antibody that binds to mouse immunoglobulin G (IgG). The biotin label allows for detection and visualization of the antibody-antigen complex.
Automatically generated - may contain errors
3 protocols using biotin labeled rabbit anti mouse igg
1
Immunohistochemical Detection of M2 Macrophages
2
Immunohistochemistry for Vascular and Macrophage Markers
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CD31 has been used as a speci c marker to identify vascular endothelial cells 11, 12 , and CD163 has been used as a speci c marker to identify M2 macrophages 7, 13, 14 . Surgically resected specimens were retrieved to perform immunohistochemistry. Sections 4 µm in thickness were depara ned and rehydrated. The sections were then subjected to endogenous peroxidase blocking in 1% H 2 O 2 solution in methanol for 15 min. Antigen retrieval was performed by autoclaving the sections at 105°C for 10 min in Dako Target Retrieval Solution (Dako, Glostrup, Denmark). Serum blocking was performed with 10% normal rabbit serum for 10 min. After H 2 O 2 and serum blocking, the slides were incubated with primary mouse monoclonal anti-CD31 antibody (1:40 dilution; Dako, Glostrup, Denmark)/anti-CD163 antibody (1:200 dilution; Leica Biosystems, Newcastle Upon Tyne, UK) at room temperature for 1 h. The secondary antibody was biotin-labeled rabbit anti-mouse IgG (1:500; Nichirei, Tokyo, Japan). Detection was performed with a DAB kit (Histo ne simple stain kit; Nichirei). The sections were counterstained with hematoxylin.
3
Immunohistochemistry for Vascular and Macrophage Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD31 has been used as a speci c marker to identify vascular endothelial cells 11, 12 , and CD163 has been used as a speci c marker to identify M2 macrophages 7, 13, 14 . Surgically resected specimens were retrieved to perform immunohistochemistry. Sections 4 µm in thickness were depara ned and rehydrated. The sections were then subjected to endogenous peroxidase blocking in 1% H 2 O 2 solution in methanol for 15 min. Antigen retrieval was performed by autoclaving the sections at 105°C for 10 min in Dako Target Retrieval Solution (Dako, Glostrup, Denmark). Serum blocking was performed with 10% normal rabbit serum for 10 min. After H 2 O 2 and serum blocking, the slides were incubated with primary mouse monoclonal anti-CD31 antibody (1:40 dilution; Dako, Glostrup, Denmark)/anti-CD163 antibody (1:200 dilution; Leica Biosystems, Newcastle Upon Tyne, UK) at room temperature for 1 h. The secondary antibody was biotin-labeled rabbit anti-mouse IgG (1:500; Nichirei, Tokyo, Japan). Detection was performed with a DAB kit (Histo ne simple stain kit; Nichirei). The sections were counterstained with hematoxylin.
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