c-Myc Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

The c-Myc protein is a transcription factor, which is encoded by the c-Myc gene on human chromosome 8q24. c-Myc is a multifunctional, nuclear phosphoprotein that functions as a transcription factor with a theoretical molecular weight of 62kDa. However, c-Myc is extremely labile and is degraded very quickly even in extracts prepared with boiling SDS sample buffer, such that a molecular weight of ~40 kDa has been observed. c-Myc is part of a heterodimeric complex with Max that acts as a potent transcriptional activator. c-Myc is modified by glycosylation and phosphorylation and has been shown to interact with numerous proteins including SMAD2, SMAD3, LSD1/KDM1A, MAD, and Sp1 (1).

A basic Helix-Loop-Helix, Leucine Zipper domain (bHLH/LZ), designated Max, specifically associates with c-Myc, N-Myc and L-Myc proteins. The Myc-Max complex binds to DNA in a sequence-specific manner under conditions where neither Max nor Myc exhibit appreciable binding. Max can also form heterodimers with other bHLH-Zip proteins, Mad and Mxi1. c-Myc plays a role in cell cycle progression, apoptosis, cellular transformation and angiogenesis (2). Mutations, overexpression, rearrangement and translocation of this gene have been associated with a variety of cancers including B-cell Lymphomas, acute myeloid leukemia, glioblastoma, stomach adenocarcinoma, and prostate adenocarcinoma (3).

References

1. Wilkinson, D. S., Tsai, W. W., Schumacher, M. A., & Barton, M. C. (2008). Chromatin-bound p53 anchors activated Smads and the mSin3A corepressor to confer transforming-growth-factor-beta-mediated transcription repression. Mol Cell Biol, 28(6), 1988-1998. doi:10.1128/mcb.01442-07

2. Pedrosa, A. R., Bodrug, N., Gomez-Escudero, J., Carter, E. P., Reynolds, L. E., Georgiou, P. N., . . . Hodivala-Dilke, K. M. (2019). Tumor Angiogenesis Is Differentially Regulated by Phosphorylation of Endothelial Cell Focal Adhesion Kinase Tyrosines-397 and -861. Cancer Res, 79(17), 4371-4386. doi:10.1158/0008-5472.Can-18-3934

3. Nagasaka, M., Tsuzuki, K., Ozeki, Y., Tokugawa, M., Ohoka, N., Inoue, Y., & Hayashi, H. (2019). Lysine-Specific Demethylase 1 (LSD1/KDM1A) Is a Novel Target Gene of c-Myc. Biol Pharm Bull, 42(3), 481-488. doi:10.1248/bpb.b18-00892

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Images	Ratings	Applications	Species	Date	Details
Enlarge	5 reviewed by: Costache Vasilescu	WB	Human	12/18/2014	View Summary Application Western Blot Sample Tested Whole cell lysate from 293T cells, in lysis buffer Species Human

Array NBL1-13414

• c-Myc Antibodies
• c-Myc ELISA Kits
• c-Myc Lysate
• c-Myc Proteins