PIK3R5 Overexpression Lysate

Western Blot: PIK3R5 Overexpression Lysate (Adult Normal) [NBL1-14424] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PIK3R5.

• Reactivity: Hu
• Applications: WB

PIK3R5 Overexpression Lysate Summary

• Description:

  PIK3R5 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC222249
  
  Accession#: NM_014308
  
  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: PIK3R5

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: 97.2 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 PIK3R5 Overexpression Lysate

• F730038I15Rik
• FOAP-2
• p101
• p101-PI3K
• Phosphatidylinositol-4,5-bisphosphate 3-kinase regulatory subunit
• phosphoinositide-3-kinase, regulatory subunit 5
• phosphoinositide-3-kinase, regulatory subunit, polypeptide p101
• PI3-kinase p101 subunit
• PI3-kinase regulatory subunit 5
• PIK3R5
• PtdIns-3-kinase regulatory subunit
• regulatory subunit 5, p101

Background

Phosphatidylinositol 3-kinase (PI 3-kinase) is composed of 85 kDa (p85) and 110 kDa (p110) subunits. p85 lacks PI 3-kinase activity and acts as an adapter, coupling p110 to activated protein tyrosine kinase. Two forms of p85 have been described (p85alpha and p85beta), each possessing one SH3 and two SH2 domains. Various p110 isoforms have been identified. p110alpha and p110beta interact with p85alpha, and p110alpha has also been shown to interact with p85beta in vitro. p110delta expression is restricted to white blood cells. It has been shown to bind p85alpha and beta, but it apparently does not phosphorylate these subunits. p110delta seems to have the capacity to autophosphorylate. p110gamma does not interact with the p85 subunits. It has been shown to be activated by alpha and betagamma heterotrimeric G proteins.

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: PIK3R5