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AB266253

Human ATP2C1 knockout HEK-293T cell line

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ATP2C1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4. 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 Images
Sanger Sequencing - Human ATP2C1 knockout HEK-293T cell line (AB266253)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP2C1 knockout HEK-293T cell line (AB266253)

Allele-1 : 1 bp deletion in exon 4

Sanger Sequencing - Human ATP2C1 knockout HEK-293T cell line (AB266253)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP2C1 knockout HEK-293T cell line (AB266253)

Allele-2 : 1 bp insertion in exon 4.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4

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?

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Properties and storage information

Gene name
ATP2C1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

ATP2C1 also known as Secretory Pathway Calcium ATPase 1 (SPCA1) is an essential calcium pump. It has a molecular mass of approximately 115 kDa. ATP2C1 is located on the Golgi membrane and participates in maintaining calcium and manganese ion homeostasis by transporting these ions from the cytosol into the Golgi apparatus. It shows expression in various tissues with high levels found in epithelial cells.
Biological function summary

The efficient functioning of ATP2C1 regulates calcium ion concentration within the Golgi impacting protein sorting and processing. This protein contributes to cellular signaling and is a part of a broader family of P-type ATPases. Although ATP2C1 does not form part of a complex its activity indirectly influences the function of other calcium-binding proteins through its ion transport role.

Pathways

ATP2C1 participates in key calcium homeostasis pathways ensuring cellular functions related to calcium-dependent processes. It plays a role in the intracellular signaling pathway that involves calcium-regulated gene expression. ATP2C1 is functionally linked to proteins such as STIM1 and Orai1 which are critical in the process of calcium reabsorption and signaling.

Dysfunctional ATP2C1 activity associates with Hailey-Hailey disease a genetic disorder causing recurring blisters and erosions of the skin. Mutations in the ATP2C1 gene result in impaired calcium ion transport contributing to the skin lesions. This condition shows links to the protein desmoglein-1 involved in cell adhesion suggesting disrupted adhesion due to altered calcium ion homeostasis.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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