Product nameHuman IDH1 knockout HeLa cell lysate
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
User storage instructions: After reconstitution, store the lysate at -80°C.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.
*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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.
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab260873 - Human IDH1 knockout HeLa cell lysate (Lyophilized) 1 x 100µg ab255552 - Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
STR AnalysisAmelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
Involvement in diseaseGlioma
Genetic variations are associated with cartilaginous tumors such as enchondroma or chondrosarcoma. Mutations of Arg-132 to Cys, Gly or His abolish the conversion of isocitrate to alpha-ketoglutarate. Instead, alpha-ketoglutarate is converted to R(-)-2-hydroxyglutarate.
Sequence similaritiesBelongs to the isocitrate and isopropylmalate dehydrogenases family.
modificationsAcetylation at Lys-374 dramatically reduces catalytic activity.
Cellular localizationCytoplasm. Peroxisome.
- Information by UniProt
- Cytosolic NADP isocitrate dehydrogenase
- Cytosolic NADP-isocitrate dehydrogenase
- Epididymis luminal protein 216
KO cell lines
Our Abpromise guarantee covers the use of ab257221 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.|
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: IDH1 knockout HeLa cell lysate (20 µg)
Lane 3: Wild-type HAP1 cell lysate (20 µg)
Lane 4: IDH1 knockout Hap1 cell lysate (20 µg)
ab230949 Anti-IDH1 antibody [EPR21002] was shown to specifically react with IDH1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264916 (knockout cell lysate ab257221) was used. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. ab230949 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) 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.
Homozygous: Insertion of the selection cassette in exon 4
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab257221 has not yet been referenced specifically in any publications.