Human HIBADH knockout HeLa cell line
- Advanced Validation
- What is this?
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Human HIBADH knockout HeLa cell line available to order. Recommended control: Human wild-type HeLa cell line (ab255928).
View Alternative Names
3 hydroxy 2 methylpropanoate:NAD(+) oxidoreductase, 3 hydroxyisobutyrate dehydrogenase mitochondrial, 3-hydroxyisobutyrate dehydrogenase, 3HIDH_HUMAN, EC 1.1.1.31, MGC40361, NS5ATP1, mitochondrial
- WB
Lab
Western blot - Human HIBADH knockout HeLa cell line (AB265888)
Lanes 1-3 : Merged signal (red and green). Green - ab175203 observed at 35 kDa. Red - loading control ab7291 observed at 50 kDa.
ab175203 Anti-HIBADH antibody [EPR12519(B)] was shown to specifically react with HIBADH in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265888 (knockout cell lysate ab257986) was used. Wild-type and HIBADH knockout samples were subjected to SDS-PAGE. ab175203 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-HIBADH antibody [EPR12519(B)] (<a href='/en-us/products/primary-antibodies/hibadh-antibody-epr12519b-ab175203'>ab175203</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
HIBADH knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human HIBADH knockout HeLa cell line (ab265888)
Lane 3:
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: 35 kDa,78 kDa,80 kDa
Observed band size: 100-125 kDa,35 kDa,80 kDa
false
- WB
Lab
Western blot - Human HIBADH knockout HeLa cell line (AB265888)
Lanes 1-3 : Merged signal (red and green). Green - ab172955 observed at 35 kDa. Red - loading control ab7291 observed at 50 kDa.
ab172955 Anti-HIBADH antibody [EPR12525(B)] was shown to specifically react with HIBADH in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265888 (knockout cell lysate ab257986) was used. Wild-type and HIBADH knockout samples were subjected to SDS-PAGE. ab172955 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-HIBADH antibody [EPR12525(B)] (<a href='/en-us/products/primary-antibodies/hibadh-antibody-epr12525b-ab172955'>ab172955</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
HIBADH knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human HIBADH knockout HeLa cell line (ab265888)
Lane 3:
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: 35 kDa
Observed band size: 35 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human HIBADH knockout HeLa cell line (AB265888)
Allele-1 : 298 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human HIBADH knockout HeLa cell line (AB265888)
Allele-2 : 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
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
This enzyme facilitates the metabolism of amino acids by converting valine-derived intermediates. HIBADH contributes significantly to energy production particularly during periods of fasting. It is not typically recognized as part of a larger protein complex functioning independently within metabolic pathways. Its role in energy homeostasis indicates its essential part in maintaining cellular function during metabolic stress.
Pathways
HIBADH is integral to the valine degradation pathway and also impacts the broader branched-chain amino acid catabolism process. These pathways are critical for recycling amino acids for ATP production. HIBADH interacts with other enzymes like acyl-CoA dehydrogenases which further process breakdown products. This interaction highlights HIBADH's connection to a consistent and efficient metabolic network ensuring effective energy yield from valine.
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
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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