Recombinant Rabbit monoclonal [H26-10] Anti-Human IgG1 H&L (Biotin) (ab201485)

Ab201485 was tested using a sandwich ELISA approach. The wells were coated with ab200699 at 5µg/ml at 50µl/well overnight at 4°C, followed by a 5% BSA blocking step for 2h RT. Human IgG1 (ab90283) was then added starting at 100 µg/ml and plasma/serum at 1:50 and gradually diluted 1:4, 50µl/well for 2h. Ab201485 was then added at 1:10,000 dilution, 50µl/well for 2h. A HRP-streptavidin (ab7403) was used at 1:10,000 dilution for 1h.

Negative IHC image of Human IgG1 H & L staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex, performed on a Leica Bond^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab201485, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of Human IgG1 H & L staining in a section of formalin-fixed paraffin-embedded normal human tonsil*, performed on a Leica Bond^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab201485, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre