Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Cyclin T1 (D1B6G) Rabbit mAb #81464

No. Size Price
81464S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
81464 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 81 Rabbit IgG
IP 1:100
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Specificity / Sensitivity

Cyclin T1 (D1B6G) Rabbit mAb recognizes endogenous levels of total cyclin T1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly642 of human cyclin T1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Cyclin T1 (D1B6G) Rabbit mAb.

IP

IP

Immunoprecipitation of cyclin T1 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cyclin T1 (D1B6G) Rabbit mAb. Western blot analysis was performed using Cyclin T1 (D1B6G) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT 116 cells and either 10 µl of Cyclin T1 (D1B6G) Rabbit 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 c-Myc Intron 1 Primers #14905, SimpleChIP® Human GAPDH Exon 1 Primers #5516, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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

Positive transcription elongation factor (P-TEFb) is a heterodimer composed of cyclin T proteins and CDK9. P-TEFb plays a critical role in the transition of the RNA polymerase II (RNAPII) machinery from transcription initiation to elongation (1). At some genes during transcription initiation, RNAPII moves approximately 50 nucleotides away from the transcription start site into the gene where it then pauses and awaits signaling for the formation of a productive transcription elongation complex (1,2). The release of this promoter proximal pausing of RNAPII is signaled by phosphorylation of the C-terminal domain (CTD) within the largest subunit of RNAPII at Ser2 of the heptapeptide repeat sequence by P-TEFb (3). This phosphorylation event is important for the recruitment of mRNA processing factors and chromatin modifiers that are necessary for proper gene expression (4,5). P-TEFb also promotes transcription elongation by phosphorylating DSIF (DRB-induced stimulating factor) and NELF (negative elongation factor), two negative elongation factors that retain RNAPII at the promoter proximal region of genes to initiate transcription elongation (6,7).

  1. Cho, S. et al. (2010) Cell Cycle 9, 1697-705.
  2. Saunders, A. et al. (2006) Nat Rev Mol Cell Biol 7, 557-67.
  3. Buratowski, S. (2009) Mol Cell 36, 541-6.
  4. Lenasi, T. and Barboric, M. (2010) RNA Biol 7, 145-50.
  5. Pirngruber, J. et al. (2009) Cell Cycle 8, 3636-42.
  6. Wada, T. et al. (1998) EMBO J 17, 7395-403.
  7. Yamada, T. et al. (2006) Mol Cell 21, 227-37.

Application References

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Protocols

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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