Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

CDK9 (C12F7) Rabbit mAb #2316

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IHC-F IF-IC F H M R Hm Mk B Dg Endogenous 42, 55 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Dg=Dog
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

CDK9 (C12F7) Rabbit mAb detects endogenous levels of total CDK9 protein, both 42 kDa and 55 kDa isoforms.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CDK9.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using CDK9 (C12F7) Rabbit mAb.

IP

IP

Immunoprecipitation of CDK9 from HeLa cells using CDK9 (C12F7) Rabbit mAb. Western blot detection was performed using the same antibody. Lane 1 is 5% input.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using CDK9 (C12F7) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded K7M2 mouse syngeneic tumor using CDK9 (C12F7) Rabbit mAb.

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen SKOV-3 xenograft using CDK9 (C12F7) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using CDK9 (C12F7) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using CDK9 (C12F7) Rabbit mAb (green). Actin filaments have been labeled with DY-555 phalloidin (red).

Background

P-TEFb is a general transcription factor that regulates transcription elongation through phosphorylation of the C-terminal tail domain (CTD) of RNA polymerase II (RNAP II). The P-TEFb complex composed of a catalytic subunit, CDK9, and its regulatory cyclin partner, which can be cyclin T1, T2a, T2b or K (reviewed in 1,2). P-TEFb is recruited by the HIV Tat protein to allow transcriptional elongation, and subsequent replication of the viral genome. Inhibition of P-TEFb function therefore has potential for HIV therapy.CDK9 exists as two isoforms, an abundant 42 kDa isoform, and a less abundant 55 kDa isoform, which contains an amino-terminal extension (3). The two forms likely have distinct purposes based on differential expression during lymphocyte activation (4,5) and on their localization within the nucleus (5).Cyclin dependent kinases (CDKs) are activated in part by cyclin binding and by phosphorylation of a conserved threonine in the T-loop domain. Phosphorylation of CDK9 at the T-loop Thr186 by an unidentified nuclear kinase may be important in P-TEFb activation (6) and regulation of HIV transcription (7). Acetylation of CDK9 at Lys44 affects its ability to phosphorylate the RNAPII CTD (8).

  1. Rice, A.P. and Herrmann, C.H. (2003) Curr HIV Res 1, 395-404.
  2. De Falco, G. and Giordano, A. Cancer Biol Ther 1, 342-7.
  3. Shore, S.M. et al. (2003) Gene 307, 175-82.
  4. Shore, S.M. et al. (2005) Gene 350, 51-8.
  5. Liu, H. and Herrmann, C.H. (2005) J Cell Physiol 203, 251-60.
  6. Chen, R. et al. (2004) J Biol Chem 279, 4153-60.
  7. Ammosova, T. et al. (2005) Retrovirology 2, 47.
  8. Fu, J. et al. (2007) Mol Cell Biol 27, 4641-51.

Application References

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Companion Products

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

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