使用敲除细胞株进行验证

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

概述

  • 产品名称
    Anti-Aconitase 2抗体[6F12BD9]
    参阅全部 Aconitase 2 一抗
  • 描述
    小鼠单克隆抗体[6F12BD9] to Aconitase 2
  • 宿主
    Mouse
  • 经测试应用
    适用于: IP, ICC/IF, IHC-Fr, WB, In-Cell ELISA, Flow Cyt, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Bovine heart mitochondria

  • 阳性对照
    • HepG2, HeLa, H9C2, and MEF cell lysates; HDFn cells and HeLa cells. Human Normal colon FFPE tissue.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

     

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

性能

应用

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.

应用 Ab评论 说明
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml. for 2 hours.
IHC-Fr 1/1000.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 85 kDa.
In-Cell ELISA Use a concentration of 1 µg/ml.
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.

靶标

图片

  • 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.

  • 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).
  • ab110321 at 1µg/ml staining Aconitase 2 in HeLa cells by Flow Cytometry (blue). Isotype control antibody (red).
  • 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

  • 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.

  • IHC-Fr image of Aconitase 2 staining on six month old rat muscle tissue using ab110321(1:1000).4% PFA fixed sections were incubated in 0.3% PBS-Tween for 1h. The sections were then incubated with ab110321 (1:1000) overnight at +4°C. The secondary antibody Alexa Fluor® 568 donkey anti-mouse was used at a 1/1000 dilution for 1h. DAPI was used at 1.43µM

    See Abreview

文献

This product has been referenced in:
  • Márquez-Jurado S  et al. Mitochondrial levels determine variability in cell death by modulating apoptotic gene expression. Nat Commun 9:389 (2018). Read more (PubMed: 29374163) »
  • Kazak L  et al. UCP1 deficiency causes brown fat respiratory chain depletion and sensitizes mitochondria to calcium overload-induced dysfunction. Proc Natl Acad Sci U S A 114:7981-7986 (2017). WB ; Mouse . Read more (PubMed: 28630339) »

See all 7 Publications for this product

客户评价及客户问答

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
Username

Abcam user community

Verified customer

提交于 Oct 10 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 May 14 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 May 11 2012

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...

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

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