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IDH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.

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Images

Western blot - Human IDH1 knockout HeLa cell line (AB264916), expandable thumbnail
  • Cell Culture - Human IDH1 knockout HeLa cell line (AB264916), expandable thumbnail
  • Sanger Sequencing - Human IDH1 knockout HeLa cell line (AB264916), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

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

Recommended products

IDH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4.

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4

Disease

Adenocarcinoma

Concentration
Loading...

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

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Western blot - Human IDH1 knockout HeLa cell line (ab264916), expandable thumbnail

    Western blot - Human IDH1 knockout HeLa cell line (ab264916)

    Lanes 1- 2: Merged signal (red and green). Green - Anti-IDH1 antibody [EPR21002] ab230949 observed at 46 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    Anti-IDH1 antibody [EPR21002] ab230949 was shown to react with IDH1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264916 (knockout cell lysate Human IDH1 knockout HeLa cell lysate ab257221) was used. Wild-type HeLa and IDH1 knockout HeLa cell lysates 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) overnight at 4°C at a 1 in 1000 dilution and a 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

  • Cell Culture - Human IDH1 knockout HeLa cell line (ab264916), expandable thumbnail

    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), expandable thumbnail

    Sanger Sequencing - Human IDH1 knockout HeLa cell line (ab264916)

    Homozygous: Insertion of the selection cassette in exon 4.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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