Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-IDH1 antibody [EPR21002] (ab230949)

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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

      see all

    • 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

    • Western blot - Anti-IDH1 antibody [EPR21002] (ab230949)
      Western blot - Anti-IDH1 antibody [EPR21002] (ab230949)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)

      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.

    • Western blot - Anti-IDH1 antibody [EPR21002] (ab230949)
      Western blot - Anti-IDH1 antibody [EPR21002] (ab230949)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR21002] (ab230949)

      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.

    • Immunoprecipitation - Anti-IDH1 antibody [EPR21002] (ab230949)
      Immunoprecipitation - Anti-IDH1 antibody [EPR21002] (ab230949)

      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 Cytometry - Anti-IDH1 antibody [EPR21002] (ab230949)
      Flow Cytometry - Anti-IDH1 antibody [EPR21002] (ab230949)

      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.

    Publishing research using ab230949? Please let us know so that we can cite the reference in this datasheet.

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