Rpb1 CTD (4H8) Mouse mAb

2629:

Immunoprecipitation , Western Blotting

Rpb1 CTD (4H8) Antibody detects endogenous levels of total Rpb1 protein (both phosphorylated and unphosphorylated forms).

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide 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 2 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 SimpleChIP^® Human RPL30 Exon 3 Primers #7014, SimpleChIP^® Human GAPDH Exon 1 Primers #5516, SimpleChIP^® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP^® Human MYT-1 Exon 1 Primers #4493. 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.

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).

1. Svejstrup, J.Q. (2002) Nat Rev Mol Cell Biol 3, 21-9.
2. Shilatifard, A. et al. (2003) Annu Rev Biochem 72, 693-715.
3. Inukai, N. et al. (2004) J Biol Chem 279, 8190-5.
4. Oelgeschläger, T. (2002) J Cell Physiol 190, 160-9.
5. Luo, Z. et al. (2001) Mutat Res 486, 259-74.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !

• 2916 CDK7 (MO1) Mouse mAb
• 4602 DNA-PK Antibody
• 9101 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody
• 9102 p44/42 MAPK (Erk1/2) Antibody
• 3936 Ubiquitin (P4D1) Mouse mAb
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9002 SimpleChIP^® Enzymatic Chromatin IP Kit (Agarose Beads)
• 9003 SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads)

For Research Use Only. Not For Use In Diagnostic Procedures.