Anti-ATPase Inhibitory Factor 1抗体[5E2D7] (ab110277)

概述

性能

应用

Our Abpromise guarantee covers the use of ab110277 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
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 10,18 kDa (predicted molecular weight: 12 kDa).
ELISA Use a concentration of 4 µg/ml. (In-Cell ELISA, 0.4 µg/well).

靶标

  • 功能Endogenous F(1)F(o)-ATPase inhibitor limiting ATP depletion when the mitochondrial membrane potential falls below a threshold and the F(1)F(o)-ATP synthase starts hydrolyzing ATP to pump protons out of the mitochondrial matrix.
  • 序列相似性Belongs to the ATPase inhibitor family.
  • 翻译后修饰Exhibits variability in chain length, mitochondria have distinct pools of protein cleaved after the 24th, 25th, and 26th amino acid.
  • 细胞定位Mitochondrion.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ATIF1_HUMAN antibody
    • ATP synthase inhibitor protein antibody
    • ATPase inhibitor antibody
    • ATPase inhibitor mitochondrial antibody
    • ATPase inhibitor protein antibody
    • ATPase inhibitory factor 1 antibody
    • ATPI antibody
    • ATPIF 1 antibody
    • Atpif1 antibody
    • ATPIP antibody
    • IF(1) antibody
    • IF1 antibody
    • Inhibitor of F(1)F(o)-ATPase antibody
    • IP antibody
    • MGC1167 antibody
    • MGC8898 antibody
    • mitochondrial antibody
    see all

Anti-ATPase Inhibitory Factor 1 antibody [5E2D7] 图像

  • ICC/IF image of ab110277 stained HepG2 cells. The cells were 4% formaldehyde fixed (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 (ab110277, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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

  • All lanes : Anti-ATPase Inhibitory Factor 1 antibody [5E2D7] (ab110277) at 1 µg/ml

    Lane 1 : Human heart mitochondria at 10 µg
    Lane 2 : Bovine heart mitochondria at 4 µg
    Lane 3 : Rat heart mitochondria at 10 µg
    Lane 4 : Mouse heart mitochondria at 10 µg


    Predicted band size : 12 kDa
  • Overlay histogram showing HepG2 cells stained with ab110277 (red line). The cells were fixed with 80% methanol (5 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 (ab110277, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab91353) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-ATPase Inhibitory Factor 1 antibody [5E2D7] (ab110277)参考文献

This product has been referenced in:

See all 7 Publications for this product

Product Wall

Please find attached a document showing a very detailed analysis of IF1 antibody. This work was done many years ago by a scientist in our laboratory. Although the datasheet does not say the antibody is IP+, the researchers found it to be IP+ and show t...

Read More
Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
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 Oct 06 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (MEF)
Specification MEF
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 Oct 06 2014

-In regards to WB and IP applications, this antibody is very useful to monitor IF1 as you can differentiate between the 10 and 18kDa band. Ratios of the two bands vary from preparation to preparation of homogenate/mitochondria even in the same species....

Read More

Thank you for contacting us.

The immunogen of this antibody does contain the transit peptide.

This antibody actually targets on an epitope that exists both in ATP- IF and COX IV subunits, which means the double bands showed on the WB ...

Read More

Thank you for your reply, and the additional information.

According to your description of the lanes in your WB, are you expected to see no bands in the 3 right lanes- due to the miRNA knout out? Please let me know.

I would also rec...

Read More

Thank you for your reply and the addtional information including the image.

The strong band at 26 kDa is probably endogenous IgG light chain -since you are using a tissue sample this could be likely. A band like this should disappear when us...

Read More

Thank you for contacting us.

I am sorry to hear that the customer sees multiple bands.
I will check with the lab regarding the bands they saw for mouse samples and let you know.

In the meantime, could you please send me the fo...

Read More

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