Overview

  • Product nameAnti-IDH3A antibody
    See all IDH3A primary antibodies
  • Description
    Rabbit polyclonal to IDH3A
  • SpecificityThis antibody reacts with IDH3A.
  • Tested applicationsSuitable for: ICC/IF, ELISA, WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Zebrafish
  • Immunogen

    A region within synthetic peptide: IPGDGIGPEI SAAVMKIFDA AKAPIQWEER NVTAIQGPGG KWMIPSEAKE, corresponding to N terminal amino acids 37-86 of Human IDH3A

  • Positive control
    • Jurkat cell lysate and human skeletal muscle tissue

Associated products

Applications

Our Abpromise guarantee covers the use of ab58641 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
ICC/IF Use a concentration of 5 µg/ml.
ELISA 1/312500.
WB Use a concentration of 1.25 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P Use a concentration of 4 - 8 µg/ml.
IP Use at an assay dependent concentration.

Target

Anti-IDH3A antibody images

  • Anti-IDH3A antibody (ab58641) at 1.25 µg/ml + Jurkat cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size : 40 kDa
    Observed band size : 40 kDa
  • All lanes : Anti-IDH3A antibody (ab58641) at 1/10000 dilution (For 16 hours at 4°C)

    Lane 1 : Mouse adipocyte whole cell lysate
    Lane 2 : Mouse adipocyte whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa


    Exposure time : 1 second

    This image is courtesy of an anonymous Abreview

    Secondary antibody - anti-rabbit HRP (ab6721)

    See Abreview

  • IDH3A was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to IDH3A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab58641.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 38kDa; IDH3A.
  • ICC/IF image of ab58641 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab58641, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human muscle tissue labelling IDH3A with ab58641 at 4µg/ml. Arrows indicate positively labelled skeletal muscle cells.

  • All lanes : Anti-IDH3A antibody (ab58641) at 2.5 µg/ml

    Lane 1 : Jurkat cell lysate
    Lane 2 : Jurkat cell lysate with blocking peptide at 2 µg/ml

    Lysates/proteins at 25 µg per lane.


    Predicted band size : 40 kDa

References for Anti-IDH3A antibody (ab58641)

This product has been referenced in:
  • Terasawa K  et al. Preferential recognition of isocitrate dehydrogenase by a rabbit monoclonal antibody (ab124797) against the C-terminal peptide of RANKL. J Immunol Methods N/A:N/A (2015). ICC/IF ; Mouse . Read more (PubMed: 25771969) »
  • Ward PS  et al. The potential for isocitrate dehydrogenase mutations to produce 2-hydroxyglutarate depends on allele specificity and subcellular compartmentalization. J Biol Chem 288:3804-15 (2013). WB ; Human . Read more (PubMed: 23264629) »

See all 5 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (adipocyte)
Loading amount 10 µg
Specification adipocyte
Gel Running Conditions Reduced Denaturing (12% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jun 11 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"