Cell Signaling Technology

Product Pathways - Apoptosis

c-Myc Antibody #9402

Applications Reactivity Sensitivity MW (kDa) Source
W IP ChIP H M R Pg Endogenous 57 to 70 Rabbit

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

Protocols

Specificity / Sensitivity

c-Myc Antibody detects endogenous levels of total c-Myc protein. This antibody is not recommended for detection of Myc-tagged fusion proteins (use Cell Signaling Technology cat. #2276 or #2278).

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues of c-Myc. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with nonspecific (control) siRNA or transfection with c-Myc siRNA. c-Myc was detected using c-Myc Antibody #9402, and p42 was detected using p42 MAPK Antibody #9108. The c-Myc Antibody confirms silencing of c-Myc expression, and the p42 MAPK Antibody is used to control for protein loading and siRNA specificity.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, BaF3 and NBT-11 cells, using c-Myc Antibody.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Daudi cells and either 10 μl of c-Myc 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 human ATF4 promoter primers, SimpleChIP® Human NPM1 Intron 1 Primers #4779, 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

Members of the Myc/Max/Mad network function as transcriptional regulators with roles in various aspects of cell behavior including proliferation, differentiation and apoptosis (1). These proteins share a common basic-helix-loop-helix leucine zipper (bHLH-ZIP) motif required for dimerization and DNA-binding. Max was originally discovered based on its ability to associate with c-Myc and found to be required for the ability of Myc to bind DNA and activate transcription (2). Subsequently, Max has been viewed as a central component of the transcriptional network, forming homodimers as well as heterodimers with other members of the Myc and Mad families (1). The association between Max and either Myc or Mad can have opposing effects on transcriptional regulation and cell behavior (1). The Mad family consists of four related proteins; Mad1, Mad2 (Mxi1), Mad3 and Mad4, and the more distantly related members of the bHLH-ZIP family, Mnt and Mga. Like Myc, the Mad proteins are tightly regulated with short half-lives. In general, Mad family members interfere with Myc-mediated processes such as proliferation, transformation and prevention of apoptosis by inhibiting transcription (3,4).

  1. Baudino, T.A. and Cleveland, J.L. (2001) Mol. Cell. Biol. 21, 691-702.
  2. Blackwood, E.M. and Eisenman, R.N. (1991) Science 251, 1211-1217.
  3. Henriksson, M. and Lüscher, B. (1996) Adv. Cancer Res. 68, 109-182.
  4. Grandori, C. et al. (2000) Annu. Rev. Cell Dev. Biol. 16, 653-699.

Application References

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Companion Products

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

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