Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

E2F-1 Antibody #3742

Applications Reactivity Sensitivity MW (kDa) Source
W ChIP H (R) Endogenous 70 Rabbit

Applications Key:  W=Western Blotting  ChIP=Chromatin IP
Reactivity Key:  H=Human  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

E2F1 Antibody detects endogenous levels of total E2F1 protein. The antibody does not cross-react with other proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy-terminal residues of human E2F1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3, HeLa, SK-N-MC and A673 cells, using E2F-1 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untransfected (-) or transfected with E2F1 siRNA (+) to silence E2F1 protein expression, using E2F-1 Antibody.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Raji cells and either 5 μl of E2F-1 Antibody 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 using SimpleChIP® Human Timeless Intron 1 Primers #7001, human DHFR promoter primers, 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

The E2F transcription factors are essential for regulation of the cell cycle (1,2). Physiological E2F is a heterodimer composed of an E2F subunit together with a DP subunit (3, 4). Six members of the E2F family have been identified, and each E2F subunit has a DNA binding and a dimerization domain. E2F-1 to -5 activate transcription. E2F-1 to -3 bind pRb, and E2F-4 and -5 bind p107 or p130, and these interactions are under cell cycle control (5-8). E2F-1 has oncogenic properties in vivo and in vitro. E2F-1 can induce apoptosis through p53-dependent and -independent mechanisms. E2F-1 is stress-responsive, and is regulated by a PI3-kinase-like kinase family such as the ATM/ATR kinases (9-11).

  1. Helin, K. (1998) Curr. Opin. Genet. Dev. 8, 28-35.
  2. Dyson, N. (1998) Genes Dev. 12, 2245-2262.
  3. Helin, K. et al. (1993) Genes Dev. 7, 1850-1861.
  4. Wu, C. et al. (1995) Mol. Cell. Biol. 15, 2536-2546.
  5. Takahashi, Y. et al. (2000) Genes Dev. 14, 804-816.
  6. Wu, L. et al. (2001) Nature 414, 457-462.
  7. Gaubatz, S. et al. (2000) Mol. Cell 6, 729-735.
  8. Hurford, R. K. et al. (1997) Genes Dev. 11, 1447-1463.
  9. Tsai, K. Y. et al. (1998) Mol. Cell 2, 293-304.
  10. Garcia, I. et al. (2000) Cell Growth Differ. 11, 91-98.
  11. Lin, W. C. et al. (2001) Genes Dev. 15, 1833-1844.

Application References

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

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