Anti-ATP synthase Immunocapture抗体[12F4AD8AF8] (ab109867)

概述

  • 产品名称Anti-ATP synthase Immunocapture抗体[12F4AD8AF8]
  • 描述
    小鼠单克隆抗体[12F4AD8AF8] to ATP synthase Immunocapture
  • 经测试应用适用于: WB, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Cow, Human
  • 免疫原

    Bovine ATP synthase

  • 阳性对照
  • 常规说明

     

    ab109867 is a sample of pure immunocapture antibody, not immobilized to a solid support.

    Product was previously marketed under the MitoSciences sub-brand.

性能

应用

Our Abpromise guarantee covers the use of ab109867 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
WB Use at an assay dependent concentration. PubMed: 22523357
Flow Cyt Use 1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
IP Use at an assay dependent concentration. 100 µg ab109867 can capture at least 50 µg ATP synthase Immunocapture (determined with 1 mg solubilized bovine heart mitochondria).

靶标

  • 相关性Complex V, also called F1F0ATPase or ATP synthase, is responsible for ATP production in oxidative phosphorylation and can work in reverse as a proton pumping ATPase. The enzyme was thought to be localized exclusively to mitochondria. However, it has recently been identified on the plasma membrane of several cell types including hepatocytes where it functions as the HDL receptor, on endothelial cells where it may act as the angiostatin receptor, and on the surface of cancer cells. The enzyme in mammals is composed of 17 subunits, five of which make up the easily detached F1. The remainder subunits are components of two stalk domains and the proton pumping F0 part of the machinery. Two of the subunits of the F0 part are encoded on mitochondrial DNA while the other subunits are nuclear encoded. Mutations in the mitochondrial-encoded subunits of ATP synthase (Complex V) cause OXPHOS disease.

Anti-ATP synthase Immunocapture antibody [12F4AD8AF8] 图像

  • Using the immunoprecipitation protocol provided, ATP synthase was isolated from various samples by antibody ab109867 crosslinked to protein G-agarose beads (ab109715). Protein bands identities were confirmed by mass spectrometry.
  • Overlay histogram showing HepG2 cells stained with ab109867 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109867, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Anti-ATP synthase Immunocapture antibody [12F4AD8AF8] (ab109867)参考文献

This product has been referenced in:
  • Xu H  et al. Uncoupling lipid metabolism from inflammation through fatty acid binding protein-dependent expression of UCP2. Mol Cell Biol 35:1055-65 (2015). Read more (PubMed: 25582199) »
  • Alavian KN  et al. An uncoupling channel within the c-subunit ring of the F1FO ATP synthase is the mitochondrial permeability transition pore. Proc Natl Acad Sci U S A 111:10580-5 (2014). WB ; Human . Read more (PubMed: 24979777) »

See all 9 Publications for this product

Product Wall

Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Mouse Tissue lysate - whole (Brain mitochondria)
Specification Brain mitochondria
Total protein in input 300 µg
Username

Abcam user community

Verified customer

提交于 Jul 18 2014


The antibody was raised against the cow purified whole enzyme to maximize affinity for native active holoenzyme.

1) Our best immunocapture/immunoprecipitation antibodies bind the native folded proteins so often do not work in Western blot...

Read More

Thank you for your reply.

I'm glad you found the information provided useful. You can use whole cell lysates with the antibody.However, it is critical to use LM as the detergent though. Other detergents such as NP40 or SDS in RIPA bufferwill ...

Read More

Thank you for contacting us and sorry for the delay in getting back to you.

I have contacted Mitosciences and they have shared the following protocols which can be used with the Anti-ATP synthase Immunocapture antibody [12F4AD8AF8] (ab109867)...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application IHC - Wholemount
Sample Fruit fly (Drosophila melanogaster) Tissue (Dissected 3rd instar larvae)
Specification Dissected 3rd instar larvae
Username

Abcam user community

Verified customer

提交于 May 01 2012

Thank you for your call today and for your interest in ab109867. Please see below for information about your testing discount code, and please let me know if you have any questions.

DISCOUNT CODE: ***
Expiration date: July 21, 2012
...

Read More

Thank you for contacting us. The immunogen was not a recombinant protein.  It was purified ATP synthase enzyme from bovine heart (the whole complex).  Hence we don’t know the exact epitope to which the protein binds.  However, we do know for certain it...

Read More

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