IDH1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
Images
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
Alternative names
Cytosolic NADP-isocitrate dehydrogenase, Epididymis luminal protein 216, Epididymis secretory protein Li 26, HEL-216, HEL-S-26, ICDH, IDCD, IDH, IDH1, IDHC_HUMAN, IDP, IDPC, Isocitrate dehydrogenase (NADP(+)) 1 cytosolic, Isocitrate dehydrogenase 1 (NADP+) soluble, Isocitrate dehydrogenase [NADP] cytoplasmic, NADP dependent isocitrate dehydrogenase cytosolic, NADP dependent isocitrate dehydrogenase peroxisomal, NADP(+)-specific ICDH, Oxalosuccinate decarboxylase, PICD
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IDH1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
IDH1
- 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
Storage
- Shipped at conditions
Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
IDH1 also known as isocitrate dehydrogenase 1 is an enzyme with a molecular weight of approximately 45 kDa. It participates in the citric acid cycle aiding the conversion of isocitrate to alpha-ketoglutarate while producing NADPH. IDH1 resides primarily in the cytoplasm and peroxisomes of cells. It finds expression in various tissues most notably the liver and brain where metabolic processes are highly active.
- Biological function summary
Isocitrate dehydrogenase 1 plays an important role in cellular metabolism by helping to maintain the NADP+/NADPH balance important for redox reactions. This enzyme operates mainly as a homodimer meaning two identical subunits form a complex for functional activity. It contributes significantly to lipid and glucose metabolism due to its production of NADPH which the cells use for biosynthetic pathways and detoxification.
- Pathways
The involvement of IDH1 in the citric acid cycle highlights its importance in energy production and metabolic regulation. This enzyme closely associates with other metabolic enzymes such as IDH2 another member of the isocitrate dehydrogenase family found in mitochondria suggesting a coordinated function between cytoplasmic and mitochondrial metabolism. Beyond its role in the citric acid cycle IDH1 also relates to the pentose phosphate pathway an alternative glucose metabolism route providing reducing equivalents and ribose 5-phosphate for nucleotide synthesis.
- Associated diseases and disorders
IDH1 mutations particularly the IDH1 R132H variant frequently appear in certain cancers like gliomas and acute myeloid leukemia (AML). These mutations lead to a neomorphic enzyme function producing 2-hydroxyglutarate instead of alpha-ketoglutarate disrupting normal cellular metabolism and aiding tumorigenesis. The mutated IDH1 also affects the TET2 protein which involves in DNA demethylation potentially altering gene expression and contributing to oncogenic pathways.
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2 product images
Western blot - Human IDH1 knockout HeLa cell lysate (ab257221)
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: IDH1 knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-IDH1 antibody [EPR21002] ab230949 observed at 46 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-IDH1 antibody [EPR21002] ab230949 Anti-IDH1 antibody [EPR21002] was shown to specifically react with IDH1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human IDH1 knockout HeLa cell line ab264916 (knockout cell lysate ab257221) was used. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-IDH1 antibody [EPR21002] ab230949 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-IDH1 antibody [EPR21002] (Anti-IDH1 antibody [EPR21002] ab230949) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: IDH1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 46 kDa
Sanger Sequencing - Human IDH1 knockout HeLa cell lysate (ab257221)
Homozygous: Insertion of the selection cassette in exon 4
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