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AB265561

Human ATP2B1 (PMCA1) knockout HeLa cell line

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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'.
2 이미지
Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell line (AB265561)
  • 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='/ko/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='/ko/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 line (AB265561)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP2B1 (PMCA1) knockout HeLa cell line (AB265561)

Homozygous : 1 bp insertion in exon 8.

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 8

Antibiotic resistance

Puromycin 1µg/mL

질병

Adenocarcinoma

Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

제품 세부 정보

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.

제품 구성

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특성 및 보관 정보

유전자명
ATP2B1
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
Zygosity
Homozygous
배송 시 보관 조건
Dry Ice
적절한 단기 보관 조건
-196°C
적절한 장기 보관 조건
-196°C

추가 정보

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.

품질 관리

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

초기 취급 지침

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

Subculture 가이드라인
  • 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.
배양 배지

DMEM (High Glucose) + 10% FBS

동결보존 배지

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

제품 프로토콜

Product promise

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