Human Isocitrate Dehydrogenase 1/IDH1 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Isocitrate Dehydrogenase 1/IDH1 in ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 843219
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human Isocitrate Dehydrogenase 1/IDH1
    Ser2-Leu414
    Accession # O75874
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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 (Catalog # 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.
Immunocytochemistry
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; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
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 (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to astrocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
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; Catalog # 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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.
  • Entrez Gene IDs:
    3417 (Human); 15926 (Mouse); 24479 (Rat)
  • Alternate Names:
    Cytosolic NADP-isocitrate dehydrogenase; EC 1.1.1.42; IDCD; IDH; IDH1; IDP; IDPC; isocitrate dehydrogenase [NADP] cytoplasmic; isocitrate dehydrogenase 1 (NADP+), soluble; Isocitrate Dehydrogenase 1; NADP(+)-specific ICDH; NADP-dependent isocitrate dehydrogenase, cytosolic; NADP-dependent isocitrate dehydrogenase, peroxisomal; Oxalosuccinate decarboxylase; PICD
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