Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line
- Advanced Validation
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ATP2B4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2.
View Alternative Names
AT2B4_HUMAN, ATP2B2, ATPase Ca++ transporting plasma membrane 4, Atp2b4, Cation transporting ATPase, DKFZp686G08106, DKFZp686M088, MXRA 1, Matrix remodelling associated 1, Matrix remodelling associated protein 1, Matrix-remodeling-associated protein 1, OTTHUMP00000034088, OTTHUMP00000034089, PMCA 4, PMCA4b, PMCA4x, Plasma membrane calcium ATPase 4, Plasma membrane calcium ATPase isoform 4, Plasma membrane calcium pump isoform 4, Plasma membrane calcium-transporting ATPase 4, Sarcolemmal calcium pump
- WB
Lab
Western blot - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line (AB264720)
Lanes 1-3 : Merged signal (red and green). Green - ab2783 observed at 140 kDa. Red - loading control ab181602 observed at 37 kDa.
ab2783 Anti-Calcium Pump PMCA4 ATPase antibody [JA9] was shown to specifically react with Calcium Pump PMCA4 ATPase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264720 (knockout cell lysate ab258321) was used. Wild-type and Calcium Pump PMCA4 ATPase knockout samples were subjected to SDS-PAGE. ab2783 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4° at 1 in 500 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (<a href='/en-us/products/primary-antibodies/calcium-pump-pmca4-atpase-antibody-ja9-ab2783'>ab2783</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ATP2B4 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line (ab264720)
Lane 3:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 138 kDa
Observed band size: 140 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line (AB264720)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line (AB264720)
Allele-1 : 1 bp deletion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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
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
PMCA4 regulates intracellular calcium levels impacting processes such as muscle contraction neurotransmitter release and cell signaling. PMCA4 does not function as part of a larger protein complex but interacts with calmodulin which modulates its activity. Calmodulin binding activates PMCA4 increasing its capability to efflux Ca2+ and maintain cellular calcium balance.
Pathways
The calcium pump PMCA4 ATPase plays a critical role in the calcium signaling pathway and the regulation of cardiac rhythm. It helps control Ca2+ concentration within the cell affecting proteins like calcineurin that respond to calcium fluctuations. PMCA4 also interrelates with the sodium-calcium exchanger integrally involved in cardiac muscle contraction and relaxation.
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
Complementary Products
Primary Antibodies
AB2783
Anti-Calcium Pump PMCA4 ATPase antibody [JA9]
BOND RX™ Validated|KO Validated
![Western blot - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (AB2783)](https://content.abcam.com/products/images/calcium-pump-pmca4-atpase-antibody-ja9-ab2783--western-blot-img10596.jpg)
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- 1
Cell Lines & Lysates
AB258321
Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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