Human HIBADH knockout HeLa cell lysate
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HIBADH KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 298 bp insertion in exon1 and Insertion of the selection cassette in exon1.
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 lysate (AB257986)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : HIBADH knockout HeLa cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
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°C 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 (<a href='/en-us/products/cell-lines/human-hibadh-knockout-hela-cell-line-ab265888'>ab265888</a>)
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 lysate (AB257986)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : HIBADH knockout HeLa cell lysate (20 μg)
Lane 3 : HepG2 cell lysate (20 μg)
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°C 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 (<a href='/en-us/products/cell-lines/human-hibadh-knockout-hela-cell-line-ab265888'>ab265888</a>)
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 lysate (AB257986)
Allele-2 : Insertion of the selection cassette in exon1
- Sanger seq
Unknown
Sanger Sequencing - Human HIBADH knockout HeLa cell lysate (AB257986)
Allele-1 : 298 bp insertion in exon1
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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
Complementary Products
Primary Antibodies
AB249857
Anti-HIBADH antibody [EPR12519(B)] - BSA and Azide free
RabMAb|Recombinant|KO Validated
![Western blot - Anti-HIBADH antibody [EPR12519(B)] - BSA and Azide free (AB249857)](https://content.abcam.com/products/images/hibadh-antibody-epr12519b-bsa-and-azide-free-ab249857--western-blot-img655.jpg)
- 0
- 0
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