BAP1 Overexpression Lysate

Western Blot: BAP1 Overexpression Lysate (Adult Normal) [NBL1-07915] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for BAP1.

• Reactivity: Hu
• Applications: WB

BAP1 Overexpression Lysate Summary

• Description:

  BAP1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200378
  
  Accession#: NM_004656
  
  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: BAP1

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: 80.2 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 BAP1 Overexpression Lysate

• BAP1
• BRCA1 associated protein-1 (ubiquitin carboxy-terminal hydrolase)
• BRCA1-associated protein 1
• Cerebral protein 6
• cerebral protein-13
• Cerebral Protein-6
• DKFZp686N04275
• EC 3.4.19.12
• FLJ35406
• FLJ37180
• hucep-6
• KIAA0272HUCEP-13
• ubiquitin carboxyl-terminal hydrolase BAP1
• ubiquitin carboxy-terminal hydrolase
• UCHL2

Background

The protein encoded by this gene localizes to the nucleus and it interacts with the RING finger domain of the breast cancer 1, early onset protein (BRCA1). This gene is thought to be a tumor suppressor gene that functions in the BRCA1 growth control pathway. There are multiple polyadenylation sites found in this gene. [provided by RefSeq].

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.

FAQs for BAP1 Lysate (NBL1-07915). (Showing 1 - 1 of 1 FAQ).

1. I just received your BAP1 Lysate. Couple of questions: 1 - Is it already in RIPA buffer? 2- If so what is the additional vial of RIPA buffer for? 3 - What is the volume of lysate in the vial? 4 - How much do you load in your gel?
   • The lysate is in RIPA bufer. The contents should be: 1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer and 1 vial of 100 µg empty vector transfected control cell lysate in RIPA buffer  and 1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 100mM DTT). The additional vial should be the empty vector control in RIPA. This is the vector without the gene cloned into it. The volume is 100ul. Use 10 ug/lane.

Bioinformatics

• Gene Symbol: BAP1