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AB265358

Human OXCT1 (SCOT) knockout HeLa cell line

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OXCT1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 1 and 1 bp insertion in exon 1.

View Alternative Names

3-oxoacid CoA-transferase 1, EC 2.8.3.5, OTTHUMP00000120012, OTTHUMP00000221550, OXCT, Oxct1, SCOT, SCOT1_HUMAN, Scot-S, Somatic-type succinyl-CoA:3-oxoacid CoA-transferase, Succinyl CoA:3 ketoacid CoA transferase, Succinyl CoA:3 ketoacid coenzyme A transferase 1 mitochondrial, Succinyl CoA:3 oxoacid CoA transferase

2 Images
Sanger Sequencing - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)
  • Sanger seq

Unknown

Sanger Sequencing - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)

Allele-2 : 1 bp insertion in exon 1.

Sanger Sequencing - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)
  • Sanger seq

Unknown

Sanger Sequencing - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)

Allele-1 : 13 bp deletion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 1 and 1 bp insertion in exon 1

Disease

Adenocarcinoma

Reactivity data

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Product details

Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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
OXCT1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

OXCT1 also known as SCOT (succinyl-CoA:3-ketoacid CoA transferase) is an enzyme that transfers CoA groups from succinyl-CoA to acetoacetate facilitating ketone body metabolism. This enzyme has a molecular mass of approximately 56 kDa. It is expressed widely in tissues with high metabolic activity including heart kidney and skeletal muscle where energy demands are significant. There OXCT1 aids in the efficient utilization of energy resources during processes like fasting and high-intensity exercise.
Biological function summary

OXCT1 functions as an important enzyme in ketolysis which is the breakdown of ketone bodies for energy production. It does not operate within a large complex but interacts closely with other enzymes involved in similar metabolic processes. OXCT1 ensures that tissues efficiently convert ketone bodies like acetoacetate and beta-hydroxybutyrate into acetyl-CoA which then feeds into the citric acid cycle to fulfill energy requirements especially when carbohydrates are sparse.

Pathways

OXCT1 is an important component of the ketone body metabolism pathway linking it to energy homeostasis and ketogenesis. It directly interacts with enzymes like acetoacetyl-CoA thiolase and beta-hydroxybutyrate dehydrogenase. Through these associations it integrates into larger metabolic pathways such as the citric acid cycle which is essential for energy production under conditions when lipids become a primary source for energy generation.

OXCT1 is implicated in conditions like ketosis-prone type 2 diabetes and mitochondrial diseases. Alterations in the enzyme's function can disrupt energy homeostasis leading to an excess of ketone bodies in the blood known as ketoacidosis particularly in diabetic patients. Additionally mutations or deficiencies in OXCT1 can be associated with disorders of mitochondrial energy metabolism with potential interactions involving proteins like pyruvate dehydrogenase and other components of the respiratory chain contributing to disease pathogenesis.

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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