Product Pathways - Cell Cycle / Checkpoint
Rpb1 CTD (4H8) Mouse mAb #2629
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP ChIP | H M R Mk (Sc) (Hm) (Dm) | Endogenous | 250 | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
ChIP=Chromatin IP
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Sc=S. cerevisiae
Hm=Hamster
Dm=D. melanogaster
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Rpb1 CTD (4H8) Antibody detects endogenous levels of total Rpb1 protein (both phosphorylated and unphosphorylated forms).
Source / Purification
Monoclonal antibodies are produced by immunizing mice with a synthetic phosphopeptide (KLH-coupled) containing 10 heptapeptide repeats [Tyr1, Ser2, Pro3, Thr4, Ser5, Pro6, Ser7] in which Ser5 is phosphorylated.
Western Blotting
Western blot analysis of extracts from various cell lines using Rpb1 CTD (4H8) Mouse mAb.
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Rpb1 CTD (4H8) Mouse mAb or 1 μl of Normal Rabbit IgG #2729 using SimpleChIP™ Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using primers specific for the transcriptionally active RPL30 and GAPDH genes and the inactive MYOD and MYT1 genes. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Background
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-β-D-ribofuranosylbenzimidazole prevents ubiquitination of Rpb1, suggesting that CTD phosphorylation is required for proteolysis (5).
- Svejstrup, J.Q. (2002) Nat. Rev. Mol. Cell. Biol. 3, 21-29.
- Shilatifard, A. et al. (2003) Annu. Rev. Biochem. 72, 693-715.
- Inukai, N. et al. (2004) J. Biol. Chem. 279, 8190-8195.
- Oelgeschlager, T. et al. (2002) J. Cell. Physiol. 190, 160-169.
- Luo, Z. et al. (2001) Mutat. Res. 486, 259-274.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
