This data was developed using ab32328, the same antibody clone in a different buffer formulation.Overlay histogram showing A431 cells stained with ab32328 (red line). The cells were fixed with 80% methanol (5 min)/ and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32328, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1?g/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

All lanes : Anti-RhoGAP antibody [EP489Y] (ab32328) at 1/500 dilution

Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : MCF-7 cell lysate
Lane 4 : K562 cell lysate
Lane 5 : A431 cell lysate
Lane 6 : 293 cell lysate
Lane 7 : NIH 3T3 cell lysate
Lane 8 : PC12 cell lysate

Predicted band size: 171 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?

This data was developed using ab32328, the same antibody clone in a different buffer formulation.

This data was developed using ab32328, the same antibody clone in a different buffer formulation.ab32328, at a 1/100 dilution, staining human RhoGAP in stomach carcinoma by Immunohistochemistry, Paraffin embedded tissue Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.