Mouse Anti-RNA Polymerase II (Rpb1 subunit, CTD, 220kD, Pol2)
RNA polymerase II is a complex of 12 proteins that participates in transcription, mRNA processing, and transcription-coupled DNA repair (1,2). The largest subunit, Rpb1, contains a unique heptapeptide: Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7 , which is repeated up to 52 times in the carboxy-terminal domain (CTD) of Rpb1 (3). This CTD is phosphorylated by cyclin-dependent kinases (cdks), p44/42 MAPK, DNA-PKcs, and c-Abl (4). DNA damage caused by UV-light, hydrogen peroxide, or cisplatin results in ubiquitination and proteasomal degradation of Rpb1 (1,3). The kinase inhibitor 5,6-dichloro-1-b-Dribofuranosylbenzimidazole prevents ubiquitination of Rpb1, suggesting that CTD phosphorylation is required for proteolysis (5).
Applications
Suitable for use in ChIP, Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution
Western Blot: 1:1000. Incubate membranes with diluted antibody, 1X TBS, 5% w/v nonfat dry milk, 0.1% Tween-20 at 4ºC with gentle shaking overnight. Immunoprecipitation: 1:50 Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic phosphopeptide containing 10 heptapeptide repeats (Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7) in which Ser5 is phosphorylated. Species Sequence Homology: Hamster, S. cerevisiae, D. melanogaster
Form
Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, <0.02% sodium azide, 50% glycerol.
Specificity
Detects endogenous levels of total Rpb1 protein (both phosphorylated and unphosphorylated forms). Species Crossreactivity: human, mouse, rat and monkey.
