Anti-Rad51L1 antibody [1 H3/13] (ab3637)

  Western blot - Rad51L1 antibody [1 H3/13] (ab3637)

Predicted band size : 42 kDa


Proteins were separated on 4–12% SDS-polyacrylamide gels and transferred onto Immun-Blot polyvinylidene difluoride membranes at 20 V for 40 minutes using a semi-dry transfer apparatus. Blots were rinsed for 5 minutes with blocking buffer (10 mm Tris-HCl (pH 8.0), 300 mm NaCl, and 0.25% Tween 20) and blocked with 15% non-fat milk in blocking buffer for 1 hour with slow rocking. Blots were incubated with primary antibody ab3637 (RAD51B) for at least 1 hour at room temperature (or overnight at 4 °C), after which they were washed with blocking buffer (3× 5 minutes) and incubated for 1 hour with horseradish peroxidase-conjugated secondary antibody. After a thorough washing with blocking buffer (6× 5 minutes), specific bands were detected by chemiluminescence on a Fuji LAS-3000 luminescent image analyzer or by exposure to x-ray film. Bands were quantified and normalized to loading controls by analyzing imported tiffs of scanned blots using ImageJ or by using the Fuji LAS-3000 software.

Image from Gildemeister OS et al, J Biol Chem. 2009 Nov 13;284(46):31945-52. Epub 2009 Sep 26, Fig S3.