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AB264916

Human IDH1 knockout HeLa cell line

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IDH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View 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

2 Images
Cell Culture - Human IDH1 knockout HeLa cell line (AB264916)
  • Cell Culture

Unknown

Cell Culture - Human IDH1 knockout HeLa cell line (AB264916)

Representative images of IDH1 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.

Sanger Sequencing - Human IDH1 knockout HeLa cell line (AB264916)
  • Sanger seq

Lab

Sanger Sequencing - Human IDH1 knockout HeLa cell line (AB264916)

Homozygous : Insertion of the selection cassette in exon 4.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

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

Reactivity data

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Product details

We will provide viable cells that proliferate on revival.

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?

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Properties and storage information

Gene name
IDH1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

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.

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.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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