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ETFA 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 ETFA knockout HEK-293T cell line (AB266513), expandable thumbnail
  • Cell Culture - Human ETFA knockout HEK-293T cell line (AB266513), expandable thumbnail
  • Sanger Sequencing - Human ETFA knockout HEK-293T cell line (AB266513), 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

Alpha-ETF, EMA, ETFA_HUMAN, Electron transfer flavoprotein alpha polypeptide, Electron transfer flavoprotein alpha subunit, Electron transfer flavoprotein subunit alpha mitochondrial, Electron transferring flavoprotein alpha polypeptide, GA2, Glutaric aciduria II

Recommended products

ETFA 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
Loading...

Properties

Gene name
ETFA
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

ETFA or Electron-Transfer-Flavoprotein alpha subunit is an essential part of the mitochondrial respiratory chain. This protein which has a molecular mass of approximately 34 kDa functions in the transfer of electrons from acyl-CoA dehydrogenases to the main respiratory chain for energy production. ETFA is commonly expressed in tissues with high-energy demands such as the liver heart and skeletal muscle. The protein forms a heterodimeric complex with its counterpart ETFB providing a critical function in electron transfer during fatty acid oxidation.

Biological function summary

ETFA operates as an important part of the electron transfer process within the mitochondria. It acts as one-half of the heterodimeric electron-transfer flavoprotein complex teaming with ETFB. This complex facilitates electron transfer from a range of acyl-CoA dehydrogenases to ETF dehydrogenase which then continues the process of electron transfer to coenzyme Q in the respiratory chain. This action is key to the breakdown of fats enabling energy extraction and processing.

Pathways

ETFA has important roles in fatty acid beta-oxidation and amino acid catabolism. It engages in these pathways by transferring electrons as mentioned interfacing with other proteins like ETF dehydrogenase. This positioning within the mitochondrial matrix enables ETFA to assist in converting fat and protein substrates into energy which the cell can use. Its molecular interactions highlight its integral position in maintaining energy homeostasis.

Associated diseases and disorders

Problems with ETFA lead to glutaric acidemia type 2 a metabolic disorder that impairs the body's ability to oxidize fatty acids and some amino acids. Deficiencies in ETFA function disrupt the electron transport to the respiratory chain causing an accumulation of intermediary metabolites. These disruptions can relate to or involve other mitochondrial components and proteins like ETFB or ETFDH. Correct diagnosis and understanding of ETFA’s role in such conditions are instrumental for targeted therapeutic approaches.

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