Human PPP2R5D knockout A-431 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human PPP2R5D knockout A-431 cell line (AB270476)
Lanes 1 - 4 : Merged signal (red and green). Green - ab188325 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab188325 was shown to react with PPP2R5D in wild-type A431 cells in western blot with loss of signal observed in PPP2R5D knockout sample. Wild-type and PPP2R5D knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188325 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PPP2R5D antibody [EPR15617-50] (<a href='/en-us/products/primary-antibodies/ppp2r5d-antibody-epr15617-50-ab188325'>ab188325</a>) at 1/10000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
PPP2R5D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PPP2R5D knockout A-431 cell line (ab270476)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 70 kDa
Observed band size: 60-65 kDa
false
- WB
Lab
Western blot - Human PPP2R5D knockout A-431 cell line (AB270476)
Lanes 1 - 4 : Merged signal (red and green). Green - ab188323 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab188323 was shown to react with PPP2R5D in wild-type A431 cells in western blot with loss of signal observed in PPP2R5D knockout sample. Wild-type and PPP2R5D knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188323 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PPP2R5D antibody [EPR15617] (<a href='/en-us/products/primary-antibodies/ppp2r5d-antibody-epr15617-ab188323'>ab188323</a>) at 1/10000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
PPP2R5D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PPP2R5D knockout A-431 cell line (ab270476)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 70 kDa
Observed band size: 60-65 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human PPP2R5D knockout A-431 cell line (AB270476)
7 bp deletion after Gly147 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human PPP2R5D knockout A-431 cell line (AB270476)
Knockout achieved by CRISPR/Cas9; X = 7 bp deletion; Frameshift = 99.91%
Complementary Products
Primary Antibodies
AB188323
Anti-PPP2R5D antibody [EPR15617]
RabMAb|Recombinant|KO Validated|Trial Size
![Immunocytochemistry/ Immunofluorescence - Anti-PPP2R5D antibody [EPR15617] (AB188323)](https://content.abcam.com/products/images/ppp2r5d-antibody-epr15617-ab188323--immunocytochemistry-immunofluorescence-img93625.jpg)
- 0
- 0
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
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
PPP2R5D plays an essential role in the regulation of cell growth and division. This protein is part of the PP2A holoenzyme complex which assembles with catalytic and scaffolding subunits. Through its regulatory influence PPP2R5D affects the phosphorylation state of several substrates thereby impacting cellular processes like cell cycle progression and apoptosis.
Pathways
PPP2R5D significantly contributes to the MAPK signaling pathway and the Wnt pathway. It modulates phosphorylation states affecting signal transduction mechanisms. In the MAPK pathway PPP2R5D interacts with proteins like ERK1/2 by influencing their activity. Similarly in the Wnt pathway it acts on components important for maintaining cellular balance and signaling fidelity.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
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