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Myogenin Overexpression Lysate

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Western Blot: Myogenin Overexpression Lysate (Adult Normal) [NBL1-13444] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Myogenin.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Myogenin Overexpression Lysate Summary

Description

Myogenin Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC208753

Accession#: NM_002479

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.
Gene
MYOG

Applications/Dilutions

Dilutions
  • Western Blot
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 (cat# 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
24.9 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.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

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.

Alternate Names for Myogenin Overexpression Lysate

  • BHLHC3
  • bHLHc3Myogenic factor-4; myogenin
  • Class C basic helix-loop-helix protein 3
  • MYF4
  • myf-4
  • MYF4myogenin
  • MYOG
  • Myogenic factor 4
  • myogenin (myogenic factor 4)
  • Myogenin

Background

Myogenin is a member of the MyoD family of myogenic basic helix-loop-helix (bHLH) transcription factors that also includes MyoD, Myf-5, and MRF4 (also known as herculinor Myf-6). MyoD family members are expressed exclusively in skeletal muscle and play a key role in activating myogenesis by binding to enhancer sequences of muscle-specific genes. The regulatory domain of MyoD is approximately 70 amino acids in length and includes both a basic DNA binding motif and a bHLH dimerization motif. MyoD family members share about 80% amino acid homology in their bHLH motifs. Transfection of myogenin and other family members into a variety of non-muscle cells has been shown to either convert these cells to myogenic cells, or to transcriptionally activate a set of otherwise unexpressed muscle-specific genes. In addition to activating muscle specific genes, members of the MyoD family members activate their own transcription and transactivate the transcription of other MyoD family members. For example, transfection of myogenin into 10T1/2 cells or Swiss 3T3 cells results in the activation of theendogenous myogenin gene as well as transactivation of MyoD. Likewise, the transfection of MyoD into these cells results in the activation of MyoD as well as the transactivation of myogenin. Each member of the MyoD family has distinct roles in muscle development; myogenin plays a key role in muscle maturation. Myogenin migrates at a molecular weight of approx. 34 kDa by SDS-PAGE.

Limitations

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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Bioinformatics

Gene Symbol MYOG