PI 3-Kinase p55 gamma Overexpression Lysate

Name: PI 3-Kinase p55 gamma Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-14422

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-14422

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for PI 3-Kinase p55 gamma Overexpression Lysate

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.

Product Specifications

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.

TMW

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.

Type

Overexpression

Scientific Data Images for PI 3-Kinase p55 gamma Overexpression Lysate

Western Blot: PI 3-Kinase p55 gamma Overexpression Lysate [NBL1-14422]

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.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: PI 3-Kinase p55 gamma

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

Long Name

PI 3-Kinase p55 gamma

Alternate Names

p55PIK, PI 3Kinase p55 gamma, PIK3R3

Gene Symbol

PIK3R3

Additional PI 3-Kinase p55 gamma Products

Product Documents for PI 3-Kinase p55 gamma Overexpression Lysate

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COA

SDS

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Product Specific Notices for PI 3-Kinase p55 gamma Overexpression Lysate

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.

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.

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