c-Myc Overexpression Lysate

Novus Biologicals | Catalog # NBL1-13414

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for c-Myc Overexpression Lysate

c-Myc Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC201611

Accession#: NM_002467

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

50.4 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for c-Myc Overexpression Lysate

Western Blot: c-Myc Overexpression Lysate [NBL1-13414]

Western Blot: c-Myc Overexpression Lysate [NBL1-13414]

Western Blot: c-Myc Overexpression Lysate (Adult Normal) [NBL1-13414] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for c-Myc.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: c-Myc

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

Long Name

v-Myc Avian Myelocytomatosis Viral Oncogene Homolog (Avian)

Alternate Names

cMyc, Myc, Myc2, Niard, Nird

Gene Symbol

MYC

Additional c-Myc Products

Product Documents for c-Myc Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for 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.

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.

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  • Name: Costache Vasilescu
    Application: Western Blot
    Sample Tested: Whole cell lysate from 293T cells, in lysis buffer
    Species: Human
    Verified Customer | Posted 12/18/2014
    WB for c-Myc in 293T cells
    c-Myc Overexpression Lysate NBL1-13414

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FAQs for c-Myc Overexpression Lysate

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  • Q: What amount of 'over-expression lysate for c-myc' did you use in this example image for the western blot?

    A: The Western blot you see on the datasheet represents the blot from a gel wherein 5ug of lysate # NBL1-13414 was loaded. However, to our customers, we generally recommend using 5-10ug of lysate because the antibody that we used in our validation of the lysate might be different from what customers would be using. Please make sure to heat the 5-10ug lysate aliquot (to be loaded) at 95-100 degrees Celsius for 5 minutes along with your other test samples before loading on to the gel.

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