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AB267249

Human COL4A3BP (CERT) knockout HEK-293T cell line

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CERT1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and 7 bp deletion 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'.
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Sanger Sequencing - Human COL4A3BP (CERT) knockout HEK-293T cell line (AB267249)
  • Sanger seq

Unknown

Sanger Sequencing - Human COL4A3BP (CERT) knockout HEK-293T cell line (AB267249)

Allele-2 : 1 bp insertion in exon 4.

Sanger Sequencing - Human COL4A3BP (CERT) knockout HEK-293T cell line (AB267249)
  • Sanger seq

Unknown

Sanger Sequencing - Human COL4A3BP (CERT) knockout HEK-293T cell line (AB267249)

Allele-1 : 7 bp deletion in exon4

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 insertion in exon 4 and 7 bp deletion 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
CERT1
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.

CERT also known as CERT protein or COL4A3BP functions in the intracellular transport of ceramide from the endoplasmic reticulum to the Golgi apparatus. CERT consists of a PH domain a middle domain and a START domain comprising a mass of approximately 68 kDa. It is expressed in various human tissues with notable expression in the brain and liver. CERT facilitates the synthesis of sphingomyelin an essential component of cellular membranes.
Biological function summary

CERT plays a central role in lipid metabolism and is part of the ceramide transfer complex. This complex regulates the balance of ceramide and sphingomyelin as these lipids influence cell signaling and apoptosis. The effective transfer of ceramide by CERT ensures proper cell function and integrity. Aberrations in CERT activity can disrupt lipid homeostasis impacting cell survival and signaling pathways.

Pathways

CERT is integrated into the sphingolipid and lipid signaling pathways. It interacts with proteins like protein kinase D which regulates its phosphorylation status and as a result its activity. In these pathways CERT's role is important for maintaining lipid composition within cell membranes ensuring the proper functioning of signaling processes that depend on lipid-mediated communication.

Disrupted CERT function associates with conditions like cancer and neurodegenerative disorders. In cancer altered ceramide levels due to CERT dysfunction can influence cell growth and survival contributing to tumor progression. For neurodegenerative disorders CERT's interaction with proteins like protein kinase C highlights its significance in maintaining neuronal cell stability and signaling disruption of which may relate to disease development.

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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