Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IDH1 antibody [EPR21002] (ab230949)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21002] to IDH1
  • Suitable for: WB, IHC-P, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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, HAP1, 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

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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 HeLa cell lysate
      Lane 2 : IDH1 knockout HeLa cell lysate
      Lane 3 : Wild-type HAP1 cell lysate
      Lane 4 : IDH1 knockout HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 46 kDa
      Observed band size: 46 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab230949 observed at 46 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab230949 Anti-IDH1 antibody [EPR21002] was shown to specifically react with IDH1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264916 (knockout cell lysate ab257221) was used. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. ab230949 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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.

       

    • 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.

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

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

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

    ab230949 has not yet been referenced specifically in any publications.

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