Human Isocitrate Dehydrogenase 1/IDH1 Antibody (843219)

R&D Systems | Catalog # MAB7049

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 843219
View all Isocitrate Dehydrogenase 1/IDH1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Isocitrate Dehydrogenase 1/IDH1
Ser2-Leu414
Accession # O75874

Specificity

Detects human Isocitrate Dehydrogenase 1/IDH1 in ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Isocitrate Dehydrogenase 1/IDH1 Antibody (843219)

Detection of Human, Mouse, and Rat Isocitrate Dehydrogenase 1/IDH1 antibody by Western Blot.

Detection of Human, Mouse, and Rat Isocitrate Dehydrogenase 1/IDH1 by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Isocitrate Dehydrogenase 1/IDH1 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Isocitrate Dehydrogenase 1/IDH1 antibody in SK-BR-3 Human Cell Line by Immunocytochemistry (ICC).

Isocitrate Dehydrogenase 1/IDH1 in SK‑BR‑3 Human Cell Line.

Isocitrate Dehydrogenase 1/IDH1 was detected in immersion fixed SK-BR-3 human breast cancer cell line using Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Isocitrate Dehydrogenase 1/IDH1 antibody in Human Brain by Immunohistochemistry (IHC-P).

Isocitrate Dehydrogenase 1/IDH1 in Human Brain.

Isocitrate Dehydrogenase 1/IDH1 was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to astrocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Isocitrate Dehydrogenase 1/IDH1 antibody in Rat Brain by Immunohistochemistry (IHC-Fr).

Isocitrate Dehydrogenase 1/IDH1 in Rat Brain.

Isocitrate Dehydrogenase 1/IDH1 was detected in perfusion fixed frozen sections of rat brain using Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to glial cell cytoplasm. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Western Blot Shows Human Isocitrate Dehydrogenase 1/IDH1 Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human Isocitrate Dehydrogenase 1/IDH1 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Isocitrate Dehydrogenase 1/IDH1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Isocitrate Dehydrogenase 1/IDH1 at approximately 46 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Isocitrate Dehydrogenase 1/IDH1 by Simple WesternTM.

Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Isocitrate Dehydrogenase 1/IDH1 at approximately 51 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human Isocitrate Dehydrogenase 1/IDH1 Antibody (843219)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed SK‑BR‑3 human breast cancer cell line

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (cortex) and perfusion fixed frozen sections of rat brain

Knockout Validated

Isocitrate Dehydrogenase 1/IDH1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Isocitrate Dehydrogenase 1/IDH1 knockout HeLa cell line.

Simple Western

10 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line

Western Blot

0.25 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and Rat‑2 rat embryonic fibroblast cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Isocitrate Dehydrogenase 1/IDH1

Isocitrate Dehydrogenase 1 (IDH1) catalyzes the oxidative decarboxylation of isocitrate to alpha ‑ketoglutarate. There are two subclasses in the IDH family, one of them utilizing NADP+ as the electron acceptor and the other using NAD+ (1). The protein encoded by this gene is the NADP+-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. In peroxisomes, IDH1 generates the NADPH required for intraperoxisomal reduction reactions. Mutations of Arg132 of human IDH1 result in a reduced ability of the enzyme to convert isocitrate to alpha ‑ketoglutarate, but the enzyme acquires the ability to generate 2-hydroxyglutarate (2HG) from  alpha ‑ketoglutarate (2). Elevated levels of the metabolite 2HG are associated with a high risk of malignant brain tumors. Arg132 mutations of IDH1 are common in high‑grade gliomas, but not in other types of tumors (3).

References

  1. Nekrutenko, A. et al. (1998) Mol. Biol. Evol. 15:1674.
  2. Dang, L. et al. (2009) Nature 462:739.
  3. Bleeker, F.E. et al. (2009) Hum. Mutat. 30:7.

Alternate Names

IDCD, IDH1, IDP, IDPC, PICD

Entrez Gene IDs

3417 (Human); 15926 (Mouse); 24479 (Rat)

Gene Symbol

IDH1

UniProt

O75874

Additional Isocitrate Dehydrogenase 1/IDH1 Products

Product Documents for Human Isocitrate Dehydrogenase 1/IDH1 Antibody (843219)

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Product Specific Notices for Human Isocitrate Dehydrogenase 1/IDH1 Antibody (843219)

For research use only

Citations for Human Isocitrate Dehydrogenase 1/IDH1 Antibody (843219)

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Protocols

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