CHIP/STUB1 Overexpression Lysate

Western Blot: CHIP Overexpression Lysate (Adult Normal) [NBP2-07696] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CHIP.

• Reactivity: Hu
• Applications: WB

CHIP/STUB1 Overexpression Lysate Summary

• Description:

  CHIP/STUB1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200310
  
  Accession#: NM_005861
  
  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: STUB1

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: 34.7 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 CHIP/STUB1 Overexpression Lysate

• Antigen NY-CO-7
• Carboxy terminus of Hsp70-interacting protein
• CHIP
• CHIPSTIP1 homology and U box-containing protein 1
• CLL-associated antigen KW-8
• E3 ubiquitin-protein ligase CHIP
• EC 6.3.2.-
• heat shock protein A binding protein 2 (c-terminal)
• HSPABP2
• NY-CO-7
• SDCCAG7
• serologically defined colon cancer antigen 7
• STIP1 homology and U-box containing protein 1
• STIP1 homology and U-box containing protein 1, E3 ubiquitin protein ligase
• STUB1
• UBOX1

Background

STUB1/CHIP is an E3 ubiquitin ligase and co-chaperone of heat-shock protein 70 (Hsp70). STUB1/CHIP interacts with Hsp70 to monitor protein folding and direct misfolded proteins for proteasomal degradation. It has been found to also associate with Hsp90 to remove phosphorylated protein tau, a protein that accumulates in the brains of Alzheimers and Down's syndrome patients. STUB1/CHIP has been reported to also have intrinsic chaperone activity and recognize non-native proteins in response to heat stress. In this way, STUB1/CHIP has been shown to be a direct chaperone for p53. Alternate names for STUB1/CHIP include carboxy terminus of Hsp70-interacting protein, STIP1 homology and U-box containing protein 1, CLL-associated antigen KW-8 antigen, serologically defined colon cancer antigen 7, heat shock protein A binding protein 2, NY-CO-7, HSPABP2, and SDCCAG7.

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: STUB1