MED6 Overexpression Lysate

Western Blot: MED6 Overexpression Lysate (Adult Normal) [NBL1-12993] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MED6.

• Reactivity: Hu
• Applications: WB

MED6 Overexpression Lysate Summary

• Description:

  MED6 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC202504
  
  Accession#: NM_005466
  
  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: MED6

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 (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: 28.2 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 MED6 Overexpression Lysate

• Activator-recruited cofactor 33 kDa component
• ARC33
• mediator complex subunit 6hMed6
• mediator of RNA polymerase II transcription subunit 6
• mediator of RNA polymerase II transcription, subunit 6 homolog (S. cerevisiae)
• mediator of RNA polymerase II transcription, subunit 6 homolog
• NY-REN-28
• Renal carcinoma antigen NY-REN-28

Background

MED6 also known as Mediator of RNA polymerase II transcription subunit 6, is a 246 amino acid protein that is 28 kDa; as a component of the Mediator complex, a co-activator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes, functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery, by direct interactions with regulatory proteins the mediator is recruited to promoters and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. Disease research is currently being performed with relation to MED6 and spondylolysis, renal carcinoma, carcinoma, renal cell carcinoma, cholesterol, and neuronitis. This protein has also been shown to have interactions with over 70 proteins including SMAD2, SMAD1, CDK8, MED15, and MED25 in developmental biology, transcriptional regulation of white adipocyte differentiation, generic transcription pathway, gene expression, and transcription ligand-dependent transcription of retinoid-target genes pathways.

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.

Bioinformatics

• Gene Symbol: MED6