Anti-SCF antibody [EP665Y] (ab52603) at 1/10000 dilution + recombinant protein (5ng)

Secondary
HRP-labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 31 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling SCF with purified ab52603 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

Immunohistochemical analysis of paraffin-embedded human kidney carcinoma tissue using ab52603 at a dilution of 1/100-1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Overlay histogram showing MCF-7 cells stained with ab52603 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52603, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min) used under the same conditions.

Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.