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ACADVL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.

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Images

Western blot - Human ACADVL (VLCAD) knockout HEK-293T cell line (AB266484), expandable thumbnail
  • Cell Culture - Human ACADVL (VLCAD) knockout HEK-293T cell line (AB266484), expandable thumbnail
  • Sanger Sequencing - Human ACADVL (VLCAD) knockout HEK-293T cell line (AB266484), expandable thumbnail

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1

Alternative names

ACAD 6, ACADV_HUMAN, Acadvl, Acyl CoA dehydrogenase very long chain, Acyl Coenzyme A dehydrogenase very long chain, LCACD, VLCAD, Very long chain specific acyl CoA dehydrogenase mitochondrial, Very long-chain specific acyl-CoA dehydrogenase, mitochondrial

Recommended products

ACADVL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.

Key facts

Cell type

HEK-293T

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1

Concentration
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Properties

Gene name

ACADVL

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

ACADVL also known as VLCAD (Very Long-Chain Acyl-CoA Dehydrogenase) is an enzyme that plays an important role in the breakdown of long-chain fatty acids within the mitochondria. It has a molecular mass of approximately 70 kDa. VLCAD is expressed in various tissues with high levels found in the heart skeletal muscle and liver. These tissues are heavily reliant on fatty acid oxidation for energy production especially when glucose availability is low.

Biological function summary

ACADVL functions within the mitochondrial matrix where it catalyzes the initial step of beta-oxidation by desaturating long-chain acyl-CoA molecules. This enzyme is a part of a larger complex that includes other mitochondrial dehydrogenases contributing to the degradation of fatty acids into acetyl-CoA units. The process is essential for energy production especially under fasting conditions or high-energy demand scenarios such as muscle contraction and cardiac function.

Pathways

VLCAD plays an essential role in the fatty acid beta-oxidation pathway. It works alongside other enzymes such as ACADM (Medium-Chain Acyl-CoA Dehydrogenase) to facilitate the sequential shortening of fatty acids which integrates into the Krebs cycle for further energy extraction. This pathway maximizes energy yield from fatty acids significantly influencing cellular metabolism and energy homeostasis.

Associated diseases and disorders

VLCAD deficiencies lead to severe metabolic conditions such as VLCAD deficiency. This disorder results in the impaired breakdown of long-chain fatty acids causing energy deficiency and the accumulation of toxic substances. Symptoms range from muscle weakness to serious cardiac complications. VLCAD deficiency's impact is often assessed in relation to proteins like ACSL1 and CPT2 which are involved in the fatty acid metabolism pathway and are central to effective energy processing in cells.

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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