Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)

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Overview

  • Product name

    Anti-Aconitase 2 antibody [EPR8283(B)]
    See all Aconitase 2 primary antibodies

  • Description

    Rabbit monoclonal [EPR8283(B)] to Aconitase 2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, IHC-Frmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Aconitase 2 aa 500-600. The exact sequence is proprietary.

  • Positive control

    • HeLa, K562, Jurkat and 293T cell lysates; Human kidney tissue.

  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab129105 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 1/50 - 1/100.
WB 1/10000 - 1/50000. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr 1/500.
  • Application notes
    Is unsuitable for IP.

    • Target

    Images

    • Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      All lanes : Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105) at 1/1000 dilution

      All lanes : Wild-type HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 85 kDa
      Observed band size: 85 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab129105 observed at 85 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab129105 was shown to specifically react with Aconitase 2 in wild-type HAP1 cells as signal was lost in Hela knockout cells. Wild-type and Hela knockout samples were subjected to SDS-PAGE. Ab129105 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      All lanes : Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105) at 1/10000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 85 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)

      ab129105, at 1/100 dilution, staining Aconitase 2 in Paraffin-embedded Human kidney tissue by immunohistochemistry.

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

    • Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Western blot - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)This image is courtesy of an abreview submitted by Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom
      All lanes : Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105) at 1/1000 dilution

      Lane 1 : 1 month old Liver Mitochondrial fraction
      Lanes 2-3 : 3 month old Liver Mitochondrial fraction
      Lane 4 : 6 month old Liver Mitochondrial fraction

      Secondary
      All lanes : Pig Polyclonal to anti-Rabbit (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 85 kDa


      Exposure time: 1 minute


      Blocking: 5% milk for 1 hour at 24°C

      See Abreview

    • Immunohistochemistry (Frozen sections) - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Immunohistochemistry (Frozen sections) - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)This image is courtesy of an abreview submitted by Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom
      IHC-Fr image of Aconitase 2 staining of six month old rat skeletal muscles using ab129105 (1:500). 4% PFA fixed sections were incubated in 0.3% PBS-Tween for 1h. The sections were then incubated with the antibody (ab129105) overnight at +4°C. The secondary antibody Alexa Fluor® 488 donkey anti-rabbit (1:1000) for 1h. DAPI was used at a concentration of 1.43µM.

      See Abreview

    • Other - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Other - Anti-Aconitase 2 antibody [EPR8283(B)] (ab129105)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Kuter K  et al. Astrocyte support is important for the compensatory potential of the nigrostriatal system neurons during early neurodegeneration. J Neurochem 148:63-79 (2019). Read more (PubMed: 30295916) »
    • Sacoman JL  et al. Mitochondrial O-GlcNAc Transferase (mOGT) Regulates Mitochondrial Structure, Function, and Survival in HeLa Cells. J Biol Chem 292:4499-4518 (2017). Read more (PubMed: 28100784) »
    See all 4 Publications for this product

    Publishing research using ab129105? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Aconitase 2 antibody [EPR8283(B)]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Liver)
    Specification
    Liver
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.3% TritonX in 0.1% PBS
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Read More

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Dec 11 2012

    Immunohistochemistry (Frozen sections) abreview for Anti-Aconitase 2 antibody [EPR8283(B)]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Skeletal muscle)
    Specification
    Skeletal muscle
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.3% TritonX in 0.1% PBS
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Read More

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted May 15 2012

    Western blot abreview for Anti-Aconitase 2 antibody [EPR8283(B)]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - other (Liver Mitochondrial fraction)
    Loading amount
    20 µg
    Specification
    Liver Mitochondrial fraction
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted May 15 2012

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