Anti-ATP5A抗体[15H4C4] - Mitochondrial Marker (ab14748)

概述

  • 产品名称Anti-ATP5A抗体[15H4C4] - Mitochondrial Marker
    参阅全部 ATP5A 一抗
  • 描述
    小鼠单克隆抗体[15H4C4] to ATP5A - Mitochondrial Marker
  • 经测试应用适用于: WB, ICC, IP, IHC-P, ICC/IF, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Monkey
  • 免疫原

    Purified mitochondrial Complex V (Cow).

  • 阳性对照
    • Human, bovine, mouse and rat heart mitochondria.
  • 常规说明

    Product was previously marketed under the MitoSciences sub-brand.

    Alternative versions available:

    Anti-ATP5A antibody (FITC) [15H4C4] - Mitochondrial Marker (ab119688)

性能

应用

Our Abpromise guarantee covers the use of ab14748 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 a concentration of 1 µg/ml. Detects a band of approximately 53 kDa.
ICC Use a concentration of 1 - 2 µg/ml.
IP Use at 5 µg/mg of lysate.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Mitochondrial membrane ATP synthase (F(1)F(0) ATP synthase or Complex V) produces ATP from ADP in the presence of a proton gradient across the membrane which is generated by electron transport complexes of the respiratory chain. F-type ATPases consist of two structural domains, F(1) - containing the extramembraneous catalytic core, and F(0) - containing the membrane proton channel, linked together by a central stalk and a peripheral stalk. During catalysis, ATP synthesis in the catalytic domain of F(1) is coupled via a rotary mechanism of the central stalk subunits to proton translocation. Subunits alpha and beta form the catalytic core in F(1). Rotation of the central stalk against the surrounding alpha(3)beta(3) subunits leads to hydrolysis of ATP in three separate catalytic sites on the beta subunits. Subunit alpha does not bear the catalytic high-affinity ATP-binding sites.
  • 组织特异性Fetal lung, heart, liver, gut and kidney. Expressed at higher levels in the fetal brain, retina and spinal cord.
  • 序列相似性Belongs to the ATPase alpha/beta chains family.
  • 翻译后修饰The N-terminus is blocked.
  • 细胞定位Mitochondrion inner membrane. Peripheral membrane protein.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ATP synthase alpha chain, mitochondrial antibody
    • ATP synthase subunit alpha antibody
    • ATP synthase subunit alpha mitochondrial antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, cardiac muscle antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, 1 antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, isoform 1, cardiac muscle antibody
    • ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit, isoform 2, non-cardiac muscle-like 2 antibody
    • ATP sythase (F1 ATPase) alpha subunit antibody
    • ATP5A antibody
    • Atp5a1 antibody
    • ATP5AL2 antibody
    • ATPA_HUMAN antibody
    • ATPM antibody
    • Epididymis secretory sperm binding protein Li 123m antibody
    • hATP1 antibody
    • HEL-S-123m antibody
    • MC5DN4 antibody
    • mitochondrial antibody
    • Mitochondrial ATP synthetase antibody
    • Mitochondrial ATP synthetase oligomycin resistant antibody
    • Modifier of Min 2 mouse homolog antibody
    • Modifier of Min 2, mouse, homolog of antibody
    • MOM2 antibody
    • OMR antibody
    • ORM antibody
    • OTTHUMP00000163475 antibody
    see all

Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker 图像

  • All lanes : Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (ab14748) at 1 µg/ml

    Lane 1 : Isolated mitochondria from human heart at 10 µg
    Lane 2 : Isolated mitochondria from bovine heart at 4 µg
    Lane 3 : Isolated mitochondria from rat heart at 10 µg
    Lane 4 : Isolated mitochondria from mouse heart at 10 µg
    Lane 5 : HepG2 lysate at 20 µg
  • ab14748 (1µg/ml) staining ATP5A in human heart (left ventricle), using an automated system (DAKO Autostainer Plus). Using this protocol there is mitochondrial staining of cardiomyocytes.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 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.
  • ab14748 staining ATP5A in MDA MB 231 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 10% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/100 in BSA + 0.02% Tween 20) for 1 hour at 21°C. A DyLight® 550-conjugated goat anti-mouse IgG polyclonal  (1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (ab14748) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 36 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab14748 stained MCF7 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 (ab14748, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • Overlay histogram showing HepG2 cells stained with ab14748 (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 (ab14748, 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 in 0.1% PBS-Tween used under the same conditions.

Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker (ab14748)参考文献

This product has been referenced in:
  • Helman A  et al. p16(Ink4a)-induced senescence of pancreatic beta cells enhances insulin secretion. Nat Med N/A:N/A (2016). IHC ; Human . Read more (PubMed: 26950362) »
  • Sen A & Cox RT Clueless is a conserved ribonucleoprotein that binds the ribosome at the mitochondrial outer membrane. Biol Open 5:195-203 (2016). Read more (PubMed: 26834020) »

See all 65 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrite buffer, pH6.0
Specification colon
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 10 2016

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Sample Human Cell (MDA MB 231 cells)
Specification MDA MB 231 cells
Permeabilization Yes - 1% Triton
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Feb 19 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (liver)
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Dec 25 2014

Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Human Cell lysate - other (mitochondrial extract from HEK293 cells)
Specification mitochondrial extract from HEK293 cells
Total protein in input 200 µg
Username

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Verified customer

提交于 Sep 23 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Brain)
Specification Brain
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (13 %)
Sample Human Cell lysate - other (Mitochondrial/cytosolic fraction from HCT116 cells)
Specification Mitochondrial/cytosolic fraction from HCT116 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

提交于 Aug 21 2013

Thank you for contacting Abcam.

Unfortunately I am not an expert of this protein and so I am not sure about whether it is mitochondrial or nuclear (I suspect nuclear), but one publication that I found when doing a literature search was:
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Thank you very much for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial of ab14748 on the order ***, which should arrive tomorrow. Please keep me updated about t...

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As discussed on phone I am happy to provide you 5% discount for purchase of any one of the discussed product + free ab123069
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