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AB258321

Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate

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ATP2B4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 1 bp insertion in exon2.
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Western blot - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate (AB258321)
  • WB

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Western blot - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate (AB258321)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : ATP2B4 knockout HeLa cell lysate (20 μg)

Lane 3 : SH-SY5Y cell lysate (20 μg)

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°C 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 3:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 138 kDa

Observed band size: 140 kDa

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Sanger Sequencing - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate (AB258321)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate (AB258321)

Allele-2 : 1 bp insertion in exon2

Sanger Sequencing - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate (AB258321)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell lysate (AB258321)

Allele-1 : 1 bp deletion in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 1 bp insertion in exon2.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB2783

Anti-Calcium Pump PMCA4 ATPase antibody [JA9]

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (AB2783)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

  • 0
  • 0
Proteins & Peptides

AB180317

Recombinant Human POLR1C protein

SDS-PAGE - Recombinant Human POLR1C protein (AB180317)

  • 0
  • 0
Cell Lines & Lysates

AB264720

Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line

Advanced Validation

Western blot - Human ATP2B4 (Calcium Pump PMCA4 ATPase) knockout HeLa cell line (AB264720)

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab258321-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ATP2B4 knockout HeLa cell lysate", "number":"AB258321-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB258321-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
ATP2B4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The Calcium Pump PMCA4 ATPase also known as ATP2B4 is a plasma membrane calcium ATPase responsible for the extrusion of Ca2+ ions from the cell. It plays an essential role in maintaining calcium homeostasis within eukaryotic cells. PMCA4 weighs approximately 134 kDa and is expressed in various tissues including the heart brain and kidney. The protein actively transports Ca2+ out of the cell using energy obtained from ATP hydrolysis.
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.

PMCA4 has been linked to conditions such as heart failure and certain neurological disorders. Abnormal expression or function of PMCA4 can dysregulate calcium levels contributing to cardiac arrhythmias. Additionally altered PMCA4 activity can affect synaptic function connecting it indirectly to conditions like schizophrenia. Its interaction with proteins like alpha-synuclein implicated in neurological disorders further highlights PMCA4's relevance in disease states.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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