Western blot analysis of phosphor-RPA2 (S33) was performed by loading whole cell lysate in 1X SDS sample buffer with 2-ME from 2 x 105 mouse embryonic fibroblast cells treated with (+) or without (-) 2mM hydroxyurea (HU) for 1.5 hours. Samples were loaded onto a 4-12 % Bis-Tris polyacrylamide gel (Product # WG1402BOX). Proteins were transferred to nitrocellulose membrane with wet/tank transfer. Membrane was blocked in 5% milk/TBST. Target was detected at approximately 35 kDa using a polyclonal anti-phospho-RPA2(S33) antibody (Product # PA5-39809) at a dilution of 1:1000 in 5% milk/TBST at 4°C overnight, followed by a secondary antibody HRP-anti-rabbit at a dilution of 1:5000 at room temperature for 1 hour. Chemiluminescent detection was performed. Data courtesy of Antibody Data Exchange Program.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic phosphopeptide derived from human RFA2 around the phosphorylation site of Ser33 (A-P-SP-Q-A)|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.4, with 50% glycerol, 150mM NaCl|
|Contains||0.02% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Required for DNA recombination, repair and replication. The activity of RP-A is mediated by single-stranded DNA binding and protein interactions. Required for the efficient recruitment of the DNA double-strand break repair factor RAD51 to chromatin in response to DNA damage.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.