Human PPP2R1B knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human PPP2R1B knockout HEK-293T cell line (AB266729)
Lanes 1-4 : Merged signal (red and green). Green - ab154815 observed at 66 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154815 Anti-PPP2R1B antibody [EPR10158] was shown to specifically react with PPP2R1B in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266729 (knockout cell lysate ab258133) was used. Wild-type and PPP2R1B knockout samples were subjected to SDS-PAGE. ab154815 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 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-PPP2R1B antibody [EPR10158] (<a href='/en-us/products/primary-antibodies/ppp2r1b-antibody-epr10158-ab154815'>ab154815</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
PPP2R1B knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human PPP2R1B knockout HEK-293T cell line (ab266729)
Lane 3:
Caco-2 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 66 kDa
Observed band size: 66 kDa
false
- Cell Culture
Lab
Cell Culture - Human PPP2R1B knockout HEK-293T cell line (AB266729)
Representative images PPP2R1B 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 a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human PPP2R1B knockout HEK-293T cell line (AB266729)
Homozygous : 37 bp insertion in exon 1
Complementary Products
![Flow Cytometry (Intracellular) - Anti-PPP2R1B antibody [EPR10158] (AB154815)](https://content.abcam.com/products/images/ppp2r1b-antibody-epr10158-ab154815--flow-cytometry-intracellular-img51180.jpg)
Reactivity data
Product details
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
PPP2R1B integrates into the PP2A holoenzyme complex a major serine/threonine phosphatase. This complex regulates numerous cellular processes including cell growth division and signal transduction. PPP2R1B plays a role in maintaining cellular homeostasis through its regulatory activities. By doing so it links to various signaling events some of which control oncogenes and tumor suppressors demonstrating the protein's influence over cell cycle regulation and apoptosis.
Pathways
PPP2R1B plays a significant role in the Raf/MEK/ERK signaling pathway and the PI3K/AKT pathway. Through these pathways PPP2R1B interacts with PDPK1 and AKT1 modulating their phosphorylation status and thereby affecting cell proliferation and survival. By influencing such pathways PPP2R1B helps determine cellular responses to external stimuli impacting processes like differentiation and metabolism that are vital for organismal function.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com