Anti-CARD15/NOD2 antibody [2D9] (ab31488) at 2 µg/ml + HEK 293 overexpressing CARD15/NOD2 at 20 µg

Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 115 kDa
Observed band size: 115 kDa


Exposure time: 150 seconds

ab31488 staining CARD15/NOD2 in human tonsil epithelium tissue section by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% H₂O₂ for 10 minutes at 25°C; antigen retrieval was by heat mediation in citrate buffer pH 6.0 for 20 minutes. Samples were incubated with primary antibody (1/150) for 20 minutes at 25°C. An undiluted HRP polymer-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody.

See Abreview

IHC image of CARD15/NOD2 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31488, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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.