Human ETFA knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ETFA KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
Western blot - Human ETFA knockout HEK-293T cell line (AB266513)
Lanes 1-2 : Merged signal (red and green). Green - ab110316 observed at 35 kDa. Red - loading control ab52901 observed at kDa.
ab110316 Anti-ETFA antibody [2B11AE8] was shown to specifically react with ETFA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266513 (knockout cell lysate ab257943) was used. Wild-type and ETFA knockout samples were subjected to SDS-PAGE. ab110316 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ETFA antibody [2B11AE8] (<a href='/en-us/products/primary-antibodies/etfa-antibody-2b11ae8-ab110316'>ab110316</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 40 µg
Lane 2:
ETFA knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 40 µg
Lane 2:
Western blot - Human ETFA knockout HEK-293T cell line (ab266513)
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 35 kDa,81 kDa
Observed band size: 35 kDa
false
- Cell Culture
Lab
Cell Culture - Human ETFA knockout HEK-293T cell line (AB266513)
Representative images ETFA knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ETFA knockout HEK-293T cell line (AB266513)
Homozygous : Insertion of the selection cassette in exon 1
Reactivity data
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?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Immunocytochemistry/ Immunofluorescence - Anti-ETFA antibody [2B11AE8] (AB110316)](https://content.abcam.com/products/images/etfa-antibody-2b11ae8-ab110316--immunocytochemistry-immunofluorescence-img114364.gif)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com