Human ACADVL (VLCAD) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
Human ACADVL (VLCAD) knockout HEK-293T cell line available to order. Recommended control: Human wild-type HEK293T cell line (ab255449).
View 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
- WB
Lab
Western blot - Human ACADVL (VLCAD) knockout HEK-293T cell line (AB266484)
Lanes 1-4 : Merged signal (red and green). Green - ab188872 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.
ab188872 Anti-ACADVL/VLCAD antibody [EPR15107(B)] was shown to specifically react with ACADVL/VLCAD in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266484 (knockout cell lysate ab257332) was used. Wild-type and ACADVL/VLCAD knockout samples were subjected to SDS-PAGE. ab188872 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ACADVL/VLCAD antibody [EPR15107(B)] (<a href='/en-us/products/primary-antibodies/acadvl-vlcad-antibody-epr15107b-ab188872'>ab188872</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ACADVL knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ACADVL (VLCAD) knockout HEK-293T cell line (ab266484)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDa
false
- Cell Culture
Unknown
Cell Culture - Human ACADVL (VLCAD) knockout HEK-293T cell line (AB266484)
Representative images of ACADVL 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 an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ACADVL (VLCAD) knockout HEK-293T cell line (AB266484)
Homozygous : Insertion of the selection cassette in exon 1
Reactivity data
Product details
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
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.
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
Product promise
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