概述

  • 产品名称
    Anti-Aconitase 2抗体
    参阅全部 Aconitase 2 一抗
  • 描述
    兔多克隆抗体to Aconitase 2
  • 经测试应用
    适用于: IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (Human) conjugated to KLH

  • 阳性对照
    • Mouse heart tissue lysate 293 cell lysate

性能

应用

Our Abpromise guarantee covers the use of ab71440 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
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/100 - 1/500. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa).

靶标

Anti-Aconitase 2 antibody 图像

  • All lanes : Anti-Aconitase 2 antibody (ab71440) at 1/60 dilution

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : 293 cell lysate

    Lysates/proteins at 35 µg per lane.


    Predicted band size : 85 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 140 kDa,20 kDa,35 kDa,48 kDa,60 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-Aconitase 2 antibody (ab71440) at 1/1000 dilution

    Lane 1 : Perfused isolated rat heart whole tissue lysate lysed with 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 0.1% SDS, 0.5% Na-deoxycholate, 1 mM Na3VO4, 20 mM NaF, 1 mM PMSF, 5 v/v % protease inhibitor cocktail
    Lane 2 : Perfused isolated rat heart whole tissue lysate lysed with T-PER Tissue Protein Extraction Reagent [# 785101; Pierce], containing 1mM Na3VO4, 20 mM NaF, 5 v/v % protease inhibitor cocktail

    Lysates/proteins at 12.5 µg per lane.


    Predicted band size : 85 kDa
    Observed band size : 85 kDa
  • Ab71440 (2µg/ml) staining Aconitase 2 in human stomach using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of Parietal cells.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ICC/IF image of ab71440 stained HepG2 cells. The cells were 4% formaldehyde (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 (ab71440, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

Anti-Aconitase 2 antibody (ab71440)参考文献

This product has been referenced in:
  • Naseri NN  et al. Novel Metabolic Abnormalities in the Tricarboxylic Acid Cycle in Peripheral Cells From Huntington's Disease Patients. PLoS One 11:e0160384 (2016). WB . Read more (PubMed: 27611087) »
  • Codazzi F  et al. Friedreich ataxia-induced pluripotent stem cell-derived neurons show a cellular phenotype that is corrected by a benzamide HDAC inhibitor. Hum Mol Genet 25:4847-4855 (2016). Read more (PubMed: 28175303) »

See all 7 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293T)
Loading amount
20 µg
Specification
HEK 293T
Gel Running Conditions
Reduced Denaturing (10% Bis Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Dec 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK 293 cells)
Total protein in input
1.5e+007 cells
Specification
HEK 293 cells
Treatment
100uM Hemin for 24 hrs and pulsed with 1mCi 35S Met and Cys for 30 min
Immuno-precipitation step
Protein G
Username

Macy Shen

Verified customer

提交于 Mar 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Muscle)
Loading amount
100 µg
Specification
Muscle
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Nov 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
50 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Oct 21 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1)
Loading amount
40 µg
Specification
THP-1
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Oct 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain)
Loading amount
160 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-12)
Blocking step
Milk 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 Dec 21 2009

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

Abcam user community

Verified customer

提交于 Nov 27 2009

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

注册