Initially quenching of endogenous peroxidase in 3% hydrogen peroxide solution followed by blocking of nonspecific binding by preincubation with Antibody Diluent with a Background Reducing Component (Dako, Glostrup, Denmark) for 20 min was performed. Then, the serial sections were stained with monoclonal mouse antichicken antibodies (SouthernBiotech, Birmingham, AL) directed against antigens, Bu-1 (clone AV20, 1:500), CD4 (clone CT-4, 1:200), CD8α (clone CT-8, 1:200), and PBS, which was used as a control, and were incubated at room temperature for 1 h. The visualization of the antigens was performed using EnVision Systems (Dako) with the 3,3′-diaminobenzidine chromogen, according to the manufacturer's instructions.
Antibody diluent with a background reducing component
Manufactured by Agilent Technologies
Sourced in Denmark
Antibody Diluent with a Background Reducing Component is a solution used to dilute antibodies for immunoassay applications. It contains a component that helps reduce non-specific background signals.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using antibody diluent with a background reducing component
1
Immunohistochemical Staining of Chicken Antigens
2
Immunohistochemical Analysis of Chicken Immune Cells
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After thawing, the sections were air-dried, and fixed in hydrogen peroxide solution to quench endogenous peroxidase, and incubate for 20 minutes with Antibody Diluent with a Background Reducing Component (Dako, Glostrup, Denmark) for blocking non-specific binding. The serial sections were incubated at room temperature for 1 hour with monoclonal mouse anti-chicken antibodies (Southern Biotech, Birmingham, AL, USA) directed against antigens: Bu-1 (clone AV20, 1:500), CD3 (clone CT-3, 1:200), CD4 (clone CT-4, 1:200), CD8α (clone CT-8, 1:200) and TCRγδ (clone TCR-1, 1:100). Anti Bu-1 antibody recognizes a monomorphic determinant on the Bu-1 B-cell associated alloantigens of both RPL6 (Bu-1a) and RPL7 (Bu-1b) lines of inbred chickens (Rothwell et al. 1996 , Igyarto et al. 2008) . The visualization of the antigens was performed using EnVision TM Systems (Dako), according to the manufacturer's instructions. All sections were counterstained with Mayer's hematoxylin.
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