Human ATP2B1 (PMCA1) knockout HeLa cell line
- Advanced Validation
- What is this?
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ATP2B1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 8. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
AT2B1_HUMAN, ATP2B1, ATPase Ca++ transporting plasma membrane 1, Plasma membrane calcium ATPase 1, Plasma membrane calcium ATPase isoform 1, Plasma membrane calcium pump isoform 1, Plasma membrane calcium-transporting ATPase 1
- WB
Lab
Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell line (AB265561)
Lanes 1-4 : Merged signal (red and green). Green - ab190355 observed at 180-245 kDa. Red - loading control ab8245 observed at 36 kDa.
ab190355 Anti-PMCA1 antibody [EPR12029] was shown to specifically react with PMCA1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265561 (knockout cell lysate ab257365) was used. Wild-type and PMCA1 knockout samples were subjected to SDS-PAGE. ab190355 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-PMCA1 antibody [EPR12029] (<a href='/en-us/products/primary-antibodies/pmca1-antibody-epr12029-ab190355'>ab190355</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ATP2B1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell line (ab265561)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
THP-1 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: 139 kDa
Observed band size: 180-245 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATP2B1 (PMCA1) knockout HeLa cell line (AB265561)
Homozygous : 1 bp insertion in exon 8.
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
PMCA1 plays a fundamental role in maintaining calcium homeostasis which is important for many cellular processes including muscle contraction nerve impulse transmission and cell signaling. It functions independently but in some contexts it may interact with other proteins to modulate its activity. The protein helps prevent toxic accumulation of calcium inside cells therefore protecting cellular integrity and function.
Pathways
PMCA1 is integral to the calcium signaling pathway where it helps regulate cellular responses to fluctuating calcium levels. It interacts with other key proteins such as calmodulin which can enhance its activity in response to increased intracellular calcium. PMCA1 also links to the phosphatidylinositol signaling pathway where its function assists in the propagation of signaling cascades that manage processes like growth and metabolism.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB190355
Anti-PMCA1 antibody [EPR12029]
BOND RX™ Validated|KO Validated|RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMCA1 antibody [EPR12029] (AB190355)](https://content.abcam.com/products/images/pmca1-antibody-epr12029-ab190355--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img115824.jpg)
- 0
- 1
Primary Antibodies
AB189524
RabMAb|Recombinant|Lab Essentials|20ul selling size|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [EPR18668] (AB189524)](https://content.abcam.com/products/images/cd44-antibody-epr18668-ab189524--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img54872.jpg)
- 5
- 1
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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