COP1 Overexpression Lysate

Western Blot: COP1 Overexpression Lysate (Adult Normal) [NBP2-06435] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for COP1.

• Reactivity: Hu
• Applications: WB

COP1 Overexpression Lysate Summary

• Description:

  COP1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210492
  
  Accession#: NM_022457
  
  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: COP1

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: 80.3 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 COP1 Overexpression Lysate

• Constitutive photomorphogenesis protein 1 homolog
• constitutive photomorphogenic protein (COP1)
• COP1RNF200FLJ10416
• E3 ubiquitin-protein ligase RFWD2
• EC 6.3.2.-
• hCOP1
• putative ubiquitin ligase COP1
• ring finger and WD repeat domain 2
• RING finger and WD repeat domain protein 2
• RING finger protein 200

Background

COP1 is an E3 ubiquitin-protein ligase that mediates ubiquitination and subsequent proteasomal degradation of target proteins. It is directly involved in p53 ubiquitination and degradation, thereby abolishing p53-dependent transcription and apoptosis. It ubiquitinates p53 independently of MDM2 or RCHY1. It probably mediates E3 ubiquitin ligase activity by functioning as the essential RING domain subunit of larger E3 complexes. In contrast, it does not constitute the catalytic RING subunit in the DCX DET1-COP1 complex that negatively regulates JUN, where the ubiquitin ligase activity is mediated by RBX1.

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: COP1