Overview

  • Product name

    Anti-IDH1 antibody [EPR21002]
    See all IDH1 primary antibodies
  • Description

    Rabbit monoclonal [EPR21002] to IDH1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human IDH1 aa 100-200. The exact sequence is proprietary.
    Database link: O75874

  • Positive control

    • WB: SH-SY5Y, Raji, C2C12, Neuro-2a, HepG2, HeLa, C6 and PC-12 whole cell lysates; Human fetal brain and fetal kidney lysates. IHC-P: Human stomach and endometrium cancer tissues; Mouse and rat kidney tissues. Flow Cyt: HeLa cells. IP: SH-SY5Y whole cell lysate.
  • General notes

     This product was previously labelled as Isocitrate dehydrogenase

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab230949 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 46 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/50.
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Involvement in disease

      Glioma
      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 similarities

      Belongs to the isocitrate and isopropylmalate dehydrogenases family.
    • Post-translational
      modifications

      Acetylation at Lys-374 dramatically reduces catalytic activity.
    • Cellular localization

      Cytoplasm. Peroxisome.
    • Information by UniProt
    • Database links

    • Alternative names

      • Cytosolic NADP isocitrate dehydrogenase antibody
      • Cytosolic NADP-isocitrate dehydrogenase antibody
      • Epididymis luminal protein 216 antibody
      • Epididymis secretory protein Li 26 antibody
      • HEL-216 antibody
      • HEL-S-26 antibody
      • ICDH antibody
      • IDCD antibody
      • IDH antibody
      • IDH1 antibody
      • IDHC_HUMAN antibody
      • IDP antibody
      • IDPC antibody
      • Isocitrate dehydrogenase (NADP(+)) 1 cytosolic antibody
      • Isocitrate dehydrogenase [NADP] cytoplasmic antibody
      • Isocitrate dehydrogenase 1 (NADP+) soluble antibody
      • NADP dependent isocitrate dehydrogenase cytosolic antibody
      • NADP dependent isocitrate dehydrogenase peroxisomal antibody
      • NADP(+)-specific ICDH antibody
      • Oxalosuccinate decarboxylase antibody
      • PICD antibody
      see all

    Images

    • All lanes : Anti-IDH1 antibody [EPR21002] (ab230949) at 1/1000 dilution

      Lane 1 : Wild type HAP1 whole cell lysate
      Lane 2 : IDH1 knockout HAP1 whole cell lysate
      Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 46 kDa


      Exposure time: 62 seconds


      ab230949 was shown to specifically react with IDH1 in wild-type HAP1 cells as signal was lost in IDH1 knockout cells. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. ab230949 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining in rat kidney (PMID:30153799). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • All lanes : Anti-IDH1 antibody [EPR21002] (ab230949) at 1/1000 dilution

      Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg
      Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
      Lane 3 : C2C12 (mouse myoblast cell line) whole cell lysate at 20 µg
      Lane 4 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 20 µg
      Lane 5 : Human fetal brain lysate at 20 µg
      Lane 6 : Human fetal kidney lysate at 20 µg
      Lane 7 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
      Lane 8 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
      Lane 9 : C6 (rat glial tumor cell line) whole cell lysate at 10 µg
      Lane 10 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 46 kDa
      Observed band size: 47 kDa
      why is the actual band size different from the predicted?



      Exposure times: Lanes 1-6: 3 minutes; Lanes 7-8: 15 seconds; Lanes 9-10: 58 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining in mouse kidney (PMID:30153799). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining in human endometrium cancer (PMID:29921847). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling IDH1 with ab230949 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nucleus staining in human stomach (PMID:27466503). Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • IDH1 was immunoprecipitated from 0.35 mg of SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate with ab230949 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230949 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: SH-SY5Y whole cell lysate 10 µg (Input). 

      Lane 2: ab230949 IP in SH-SY5Y whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230949 in SH-SY5Y whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 30 seconds.

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling IDH1 with ab230949 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    References

    ab230949 has not yet been referenced specifically in any publications.

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