p73 Overexpression Lysate

Western Blot: p73 Overexpression Lysate (Adult Normal) [NBL1-17209] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for p73.

• Reactivity: Hu
• Applications: WB

p73 Overexpression Lysate Summary

• Description:

  p73 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC220864
  
  Accession#: NM_005427
  
  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: TP73

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: 69.4 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 p73 Overexpression Lysate

• p53-related protein
• P73p53-like transcription factor
• TA p73
• TAp73
• tumor protein p73

Background

p73 is a transcription factor that shares a significant amino acid identity with p53 and p63 (1). p73 and p63 can both activate p53-responsive genes and induce apoptosis upon over expression (2) Unlike p53, p73 is not induced by DNA damage and is not a target for inactivation by viral oncoproteins (3). Similar to p63, p73 has an additional coding region in the C-terminus (not found in p53) that can undergo complex alternative splicing giving rise to 3 splice forms (alpha, beta, and gamma) (4). Activation of p73 requires the presence of functional c-Abl. It has also be found that mutant p53 can bind to and inactivate p73 (5).

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: TP73