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AB257365

Human ATP2B1 (PMCA1) knockout HeLa cell lysate

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ATP2B1 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 insertion in exon8.
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Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell lysate (AB257365)
  • WB

Lab

Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell lysate (AB257365)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : ATP2B1 knockout HeLa cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : THP-1 cell lysate (20 ug)

ab190355 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 (<a href='/en-us/products/cell-lines/human-atp2b1-pmca1-knockout-hela-cell-line-ab265561'>ab265561</a>)

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 Sequencing - Human ATP2B1 (PMCA1) knockout HeLa cell lysate (AB257365)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP2B1 (PMCA1) knockout HeLa cell lysate (AB257365)

Homozygous : 1 bp insertion in exon8

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 insertion in exon8.

Disease

Adenocarcinoma

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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)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

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

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Proteins & Peptides

AB288799

Recombinant Human VCAM1 protein (Active)

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Biological Activity - Recombinant Human VCAM1 protein (Active) (AB288799)

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Cell Lines & Lysates

AB265561

Human ATP2B1 (PMCA1) knockout HeLa cell line

Advanced Validation

Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell line (AB265561)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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":"ab257365-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ATP2B1 knockout HeLa cell lysate", "number":"AB257365-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257365-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
ATP2B1
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 plasma membrane calcium ATPase 1 (PMCA1) also known by its gene name ATP2B1 is an essential protein responsible for the regulation of intracellular calcium levels. Mechanically PMCA1 functions as a calcium pump actively transporting calcium ions out of the cell using ATP hydrolysis. This protein has a molecular mass of approximately 135 kDa. PMCA1 is widely expressed in many tissues but shows higher expression in excitable cells such as neurons and muscle cells where rapid calcium signaling is critical.
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.

PMCA1 has associations with conditions impacting calcium regulation such as hypertension and cardiovascular diseases. Dysfunctional regulation of PMCA1 can lead to abnormal calcium homeostasis contributing to such disorders. Furthermore PMCA1's interaction with proteins like the sodium-calcium exchanger (NCX) suggests a complex regulatory network that when disrupted may trigger or exacerbate 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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