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Validated using a knockout cell line

Anti-Aconitase 2 antibody [6F12BD9] (ab110321)

Reviews (4)Q&A (1)References (26)
Western blot - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
  • Immunocytochemistry/ Immunofluorescence - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
  • Flow Cytometry - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
  • Western blot - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)

Key features and details

  • Mouse monoclonal [6F12BD9] to Aconitase 2
  • Suitable for: ICC/IF, WB, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647

Overview

  • Product name

    Anti-Aconitase 2 antibody [6F12BD9]
    See all Aconitase 2 primary antibodies

  • Description

    Mouse monoclonal [6F12BD9] to Aconitase 2

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • HepG2, HeLa, H9C2, and MEF cell lysates; HDFn cells and HeLa cells. Human Normal colon FFPE tissue.

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab110321 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 1 µg/ml.

for 2 hours.
WB (3)
Use a concentration of 1 µg/ml. Predicted molecular weight: 85 kDa.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 5 µg/ml.
Notes
ICC/IF
Use a concentration of 1 µg/ml.

for 2 hours.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 85 kDa.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 5 µg/ml.

Target

Images

  • Western blot - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    Western blot - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Aconitase 2 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)                

    Lane 4: Jurkat whole cell lysate (20 µg)

     

    Lanes 1 - 4: Merged signal (red and green). Green - ab110321 observed at 90 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab110321 was shown to specifically react with Aconitase 2 in wild-type HAP1 cells as signal was lost in Aconitase 2 knockout cells. Wild-type and Aconitase 2 knockout samples were subjected to SDS-PAGE. ab110321 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    Immunocytochemistry/ Immunofluorescence - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    ab110321 at 1µg/ml staining Aconitase 2 in HDFn cells by Immunocytochemistry (4% paraformaldehyde fixed and 0.1% Triton X-100 permeabilized) followed by Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour (red).
  • Flow Cytometry - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    Flow Cytometry - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    ab110321 at 1µg/ml staining Aconitase 2 in HeLa cells by Flow Cytometry (blue). Isotype control antibody (red).
  • Western blot - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    Western blot - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    All lanes : Anti-Aconitase 2 antibody [6F12BD9] (ab110321) at 1 µg/ml

    Lane 1 : HepG2 (human)
    Lane 2 : HeLa (human)
    Lane 3 : H9C2 (rat cardiomyocyte)
    Lane 4 : MEF (mouse fibroblast)

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 85 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aconitase 2 antibody [6F12BD9] (ab110321)

    IHC image of Aconitase 2 staining in Human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110321, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References (26)

Publishing research using ab110321? Please let us know so that we can cite the reference in this datasheet.

ab110321 has been referenced in 26 publications.

  • Jing X  et al. The detrimental effect of iron on OA chondrocytes: Importance of pro-inflammatory cytokines induced iron influx and oxidative stress. J Cell Mol Med 25:5671-5680 (2021). PubMed: 33942503
  • Charif M  et al. Dominant ACO2 mutations are a frequent cause of isolated optic atrophy. Brain Commun 3:fcab063 (2021). PubMed: 34056600
  • Bharath LP  et al. Metformin Enhances Autophagy and Normalizes Mitochondrial Function to Alleviate Aging-Associated Inflammation. Cell Metab 32:44-55.e6 (2020). PubMed: 32402267
  • Sheikh MH  et al. Immuno-metabolic impact of the multiple sclerosis patients' sera on endothelial cells of the blood-brain barrier. J Neuroinflammation 17:153 (2020). PubMed: 32386505
  • Hamilton J  et al. Mutant huntingtin does not cross the mitochondrial outer membrane. Hum Mol Genet 29:2962-2975 (2020). PubMed: 32821928
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Western blot abreview for Anti-Aconitase 2 antibody [6F12BD9]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Mitochondria isolated from mouse brain)
Gel Running Conditions
Reduced Denaturing (4-12% SDS-PAGE, Laemmli buffer with 10% β-Mercaptoethanol, 10 min at +55C denaturation. Immobilon-FL transfer membranes (Millipore), overnight transfer at 4C.)
Loading amount
15 µg
Specification
Mitochondria isolated from mouse brain
Blocking step
SEA Block Blocking Buffer as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 17 2018

Western blot abreview for Anti-Aconitase 2 antibody [6F12BD9]

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (12% tris-glycine gel)
Sample
Human Tissue lysate - whole (heart)
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 10 2014

Western blot abreview for Anti-Aconitase 2 antibody [6F12BD9]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Brain mitochondria lysate)
Loading amount
2 µg
Specification
Brain mitochondria lysate
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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 14 2012

Immunohistochemistry (Frozen sections) abreview for Anti-Aconitase 2 antibody [6F12BD9]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Skeletal muscles)
Specification
Skeletal muscles
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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 11 2012

Question

Hello, I'm on the search for an antibody against aconitase2 that can be used for IP in MEF cells, and I see that you have two possible candidates, ab110321 and ab110320. I'm wondering if there's any major differences between these two, and which you would recommend for my use. Thanks so much. Sincerely,

Read More
Answer

Thank you for contacting us. Both ACO2 antibodies cross-react with mouse, however ab110320 is more suitable for immunoprecipitation. Ab110321 is a good WB antibody. In this particular case, I would recommend using ab110320 then. I hope this information is helpful. Please do not hesitate to contact us if you need any more advice or information.

Read More

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