Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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PIK3R2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
View Alternative Names
P85B_HUMAN, PI3 kinase p85 beta subunit, PI3 kinase p85 subunit beta, PI3-kinase regulatory subunit beta, PI3-kinase subunit p85-beta, PI3K, PI3K regulatory subunit beta, PIK3R 2, Phosphatidylinositol 3 kinase, Phosphatidylinositol 3 kinase regulatory beta subunit, Phosphatidylinositol 3 kinase regulatory subunit polypeptide 2, Phosphatidylinositol 3 kinase, regulatory subunit, polypeptide 2 (p85 beta), Phosphatidylinositol 3-kinase 85 kDa regulatory subunit beta, Phosphatidylinositol 3-kinase regulatory subunit beta, Phosphoinositide 3 kinase regulatory subunit 2, Phosphoinositide 3 kinase regulatory subunit 2 (beta), Phosphoinositide 3 kinase regulatory subunit polypeptide 2, Phosphoinositide 3 kinase regulatory subunit polypeptide 2 (p85 beta), Phosphoinositide 3 kinase, regulatory subunit 2 (p85 beta), PtdIns 3 kinase p85 beta, PtdIns-3-kinase regulatory subunit beta, PtdIns-3-kinase regulatory subunit p85-beta, p85, p85 beta
- WB
Unknown
Western blot - Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (AB266799)
Lanes 1- 2 : Merged signal (red and green). Green - ab28356 observed at 85 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab28356 was shown to react with PI 3 Kinase p85 beta in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266799 (knockout cell lysate ab257586) was used. Wild-type HEK-293T and PIK3R2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab28356 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PI 3 Kinase p85 beta antibody [T15] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-beta-antibody-t15-ab28356'>ab28356</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PIK3R2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (ab266799)
Predicted band size: 81 kDa
Observed band size: 85 kDa
false
- WB
Lab
Western blot - Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (AB266799)
Lanes 1- 2 : Merged signal (red and green). Green - ab180967 observed at 85 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab180967 was shown to react with PI 3 Kinase p85 beta in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266799 (knockout cell lysate ab257586) was used. Wild-type HEK-293T and PIK3R2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab180967 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-beta-antibody-epr18416-ab180967'>ab180967</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PIK3R2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (ab266799)
Predicted band size: 56 kDa,81 kDa
Observed band size: 56 kDa,85 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (AB266799)
Homozygous : 1 bp insertion in exon 2
- Cell Culture
Unknown
Cell Culture - Human PIK3R2 (PI 3 Kinase p85 beta) knockout HEK-293T cell line (AB266799)
Representative images of PIK3R2 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
The regulatory subunit PI 3-kinase p85 beta is responsible for maintaining the balance of PI3K activity within the cellular environment. It forms a dynamic complex with the catalytic subunit influencing downstream signaling pathways. The presence of p85 beta ensures the stability and function of the PI3K complex essential for processes such as cell growth survival and metabolism.
Pathways
This protein plays a significant role in the PI3K/AKT/mTOR signaling pathway and also contributes to insulin signaling. It partners with p110 and mediates the transmission of signals from receptor tyrosine kinases (RTKs) like insulin receptors. The regulation by p85 beta affects protein synthesis and glucose uptake highlighting its function in cellular energy management.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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