Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-IDH2 antibody [EPR7576] - BSA and Azide free (ab246343)

Overview

  • Product name

    Anti-IDH2 antibody [EPR7576] - BSA and Azide free
    See all IDH2 primary antibodies
  • Description

    Rabbit monoclonal [EPR7576] to IDH2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human IDH2 aa 50-150. The exact sequence is proprietary.

  • Positive control

    • Human thyroid gland carcinoma tissue; Molt-4, K562, 293T and HepG2 whole cell lysate (ab7900).
  • General notes

    Ab246343 is the carrier-free version of ab129180. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab246343 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab246343 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 Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ICC/IF or IP.
  • Target

    • Function

      Plays a role in intermediary metabolism and energy production. It may tightly associate or interact with the pyruvate dehydrogenase complex.
    • Involvement in disease

      D-2-hydroxyglutaric aciduria 2
      Glioma
      enetic variations are associated with cartilaginous tumors such as enchondroma or chondrosarcoma.
    • Sequence similarities

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

      Acetylation at Lys-413 dramatically reduces catalytic activity. Deacetylated by SIRT3.
    • Cellular localization

      Mitochondrion.
    • Information by UniProt
    • Database links

    • Alternative names

      • D2HGA2 antibody
      • ICD-M antibody
      • IDH antibody
      • IDH2 antibody
      • IDHM antibody
      • IDHP_HUMAN antibody
      • IDP antibody
      • IDPM antibody
      • Isocitrate dehydrogenase [NADP], mitochondrial antibody
      • Isocitrate dehydrogenase 2 (NADP+), mitochondrial antibody
      • mNADP-IDH antibody
      • NADP(+)-specific ICDH antibody
      • Oxalosuccinate decarboxylase antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IDH2 knockout HAP1 cell lysate (20 µg) 
      Lane 3: K562 cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg) 
      Lanes 1 - 4: Merged signal (red and green). Green - ab129180 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab129180 was shown to recognize IDH2 when IDH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and IDH2 knockout samples were subjected to SDS-PAGE. ab129180 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129180).

    • ab129180, at 1/250 dilution, staining IDH2 in Formalin-fixed, Paraffin-embedded Human thyroid gland carcinoma by Immunohistochemistry.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129180).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Overlay histogram showing MCF7 cells stained with ab129180 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab129180, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129180).

    References

    ab246343 has not yet been referenced specifically in any publications.

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