PI 3-Kinase p55 gamma Overexpression Lysate

Western Blot: PIK3R3 Overexpression Lysate (Adult Normal) [NBL1-14422] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PIK3R3.

• Reactivity: Hu
• Applications: WB

PI 3-Kinase p55 gamma Overexpression Lysate Summary

• Description:

  PI 3-Kinase p55 gamma Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC202039
  
  Accession#: NM_003629
  
  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: PIK3R3

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: 54.3 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 PI 3-Kinase p55 gamma Overexpression Lysate

• DKFZp686P05226
• FLJ41892
• p55
• p55-GAMMA
• p55PIK
• Phosphatidylinositol 3-kinase 55 kDa regulatory subunit gamma
• phosphatidylinositol 3-kinase regulatory subunit gamma
• phosphatidylinositol 3-kinase, regulatory subunit, polypeptide 3 (p55, gamma)
• phosphoinositide-3-kinase, regulatory subunit 3 (gamma)
• PI 3Kinase p55 gamma
• PI 3-Kinase p55 gamma
• PI3K regulatory subunit gamma
• PI3-kinase regulatory subunit gamma
• PI3-kinase subunit p55-gamma
• PIK3R3
• PtdIns-3-kinase regulatory subunit gamma
• ptdIns-3-kinase regulatory subunit p55-gamma
• regulatory subunit, polypeptide 3 (p55, gamma)

Background

PI 3-Kinase p55 gamma, also referred to as PI3Kp55 gamma or p55gamma, is a regulatory subunit of phosphatidylinositol 3-kinase (PI 3-Kinase / PI3K) (1). Class 1A PI 3-Kinases are heterodimers consisting of one 110 kDa catalytic subunit (p110 alpha, beta, or delta) and one regulatory subunit (p85 alpha, p85 beta, p55 alpha, p50 alpha, or p55 gamma) (1). The p55 gamma regulatory subunit is encoded by the PIK3R3 gene and is predominantly expressed in the brain (2). Human PI 3 Kinase p55 gamma protein is 461 amino acids (aa) in length with a theoretical molecular weight of ~54 kDa (3). Structurally, PI 3-Kinase p55 gamma contains a proline rich domain (aa 34 - 44) and two Src homology domains (SH2, aa 65 - 160 and 358 - 452) (1,3). In general, Class 1A PI 3-Kinases are activated through the binding of membrane-bound tyrosine kinase receptors, such as insulin-like growth factor 1 (IGF-1) (1,2,4). Activation of PI 3-Kinase results in the phosphorylation of phosphatidyl inositol and a downstream signaling cascade of the AKT/mTOR pathway (2,4). PI 3-Kinase pathway activation results in a number of cellular responses including proliferation, survival, growth, migration, membrane trafficking, and metabolism (2). The PI3-Kinase/AKT/mTOR pathway has been shown to be dysregulated in a number of cancers, largely through loss or inactivation of the tumor suppressor PTEN (4). Furthermore, one study found that anaplastic lymphoma kinase (ALK) promoted cell migration during brain development specifically through the p55 gamma regulatory subunit of PI 3-Kinase (5).

References

1. Backer J. M. (2010). The regulation of class IA PI 3-kinases by inter-subunit interactions. Current Topics in Microbiology and Immunology. https://doi.org/10.1007/82_2010_52

2. Hirsch, E., Costa, C., & Ciraolo, E. (2007). Phosphoinositide 3-kinases as a common platform for multi-hormone signaling. The Journal of Endocrinology. https://doi.org/10.1677/JOE-07-0097

3. Uniprot (Q92569)

4. Yang, J., Nie, J., Ma, X., Wei, Y., Peng, Y., & Wei, X. (2019). Targeting PI3K in cancer: mechanisms and advances in clinical trials. Molecular Cancer. https://doi.org/10.1186/s12943-019-0954-x

5. Seo, M., Kim, J. H., & Suk, K. (2017). Role of the p55-gamma subunit of PI3K in ALK-induced cell migration: RNAi-based selection of cell migration regulators. Cell Adhesion & Migration. https://doi.org/10.1080/19336918.2016.1202385

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: PIK3R3