#13987 c-Myc/N-Myc (D3N8F) Rabbit mAb
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|c-Myc/N-Myc (D3N8F) Rabbit mAb recognizes endogenous levels of total c-Myc and N-Myc proteins.|
|Monoclonal antibody is produced by immunizing animals with recombinant protein specific to a central region within human c-Myc protein.|
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Western blot analysis of extracts from various cell lines using c-Myc/N-Myc (D3N8F) Rabbit mAb.
Western blot analysis of extracts from HT-29 cells, untreated (-) or treated with JQ1 (1 μM; 72 hr; +), using c-Myc/N-Myc (D3N8F) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, the BET bromodomain inhibitor JQ1 inhibits c-Myc expression.
Western blot analysis of extracts from K562 [c-Myc (+) / N-Myc (-)] and IMR-32 [c-Myc (-) / N-Myc (+)] cell lines using cMyc/N-Myc (D3N8F) Rabbit mAb (left), N-Myc (D1V2A) Rabbit mAb #84406 (middle), or β-Actin (13E5) Rabbit mAb #4970 (right).
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® c-Myc siRNA I #6341 (+), using c-Myc/N-Myc (D3N8F) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The c-Myc/N-Myc (D3N8F) Rabbit mAb confirms silencing of c-Myc expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
Confocal immunofluorescent analysis of HT-29 cells, untreated (left) or treated with bromodomain inhibitor JQ1 (1 μM, 72 hr; right), using c-Myc/N-Myc (D3N8F) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Flow cytometric analysis of HT29 cells using c-Myc/N-Myc (D3N8F) Rabbit mAb, treated with JQ1 (1uM, 72 hours at 37C) (blue) and untreated (green). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Daudi cells and c-Myc/N-Myc (D3N8F) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across NPM1, a known target gene of c-Myc (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Daudi cells and either c-Myc/N-Myc (D3N8F) Rabbit mAb or 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.
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).
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SignalSilence is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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