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

製品の概要

  • 製品名
    Anti-ATP5A antibody [15H4C4] - Mitochondrial Marker
    ATP5A 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [15H4C4] to ATP5A - Mitochondrial Marker
  • アプリケーション
    適用あり: WB, ICC, IP, IHC-P, ICC/IF, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Monkey
  • 免疫原

    Purified mitochondrial Complex V (Cow).

  • ポジティブ・コントロール
    • Human, bovine, mouse and rat heart mitochondria.
  • 特記事項

    This antibody clone [15H4C4] is manufactured by Abcam.
    We have the following conjugates available:

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

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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 1 - 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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    Fetal lung, heart, liver, gut and kidney. Expressed at higher levels in the fetal brain, retina and spinal cord.
  • 配列類似性
    Belongs to the ATPase alpha/beta chains family.
  • 翻訳後修飾
    The N-terminus is blocked.
  • 細胞内局在
    Mitochondrion inner membrane. Peripheral membrane protein.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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 画像

  • ICC/IF image of ab14748 stained HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab14748 at 1ug/ml (shown in red) and ab6160 (Rat monoclonal to Tubulin) at 1µg/ml (shown in green). This was followed by an incubation at room temperature for 1h with ab150119, Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed, at 0.5ug/ml (shown in red) and ab150165, Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed, at 0.5ug/ml (shown in green).

    Nuclear DNA was labelled with DAPI (shown in blue).

     

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

  • 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) 使用論文

This product has been referenced in:
  • Schatton D  et al. CLUH regulates mitochondrial metabolism by controlling translation and decay of target mRNAs. J Cell Biol 216:675-693 (2017). WB ; Mouse . Read more (PubMed: 28188211) »
  • Gómez-Serrano M  et al. Differential proteomic and oxidative profiles unveil dysfunctional protein import to adipocyte mitochondria in obesity-associated aging and diabetes. Redox Biol 11:415-428 (2017). WB ; Human . Read more (PubMed: 28064117) »

See all 120 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (AML12 hepatocytes, mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Treatment
18
Specification
AML12 hepatocytes, mitochondrial fraction
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Verified customer

投稿 Sep 29 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
American Robin (Turdis migratorius) Cell lysate - other (fibroblasts, mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing (18)
Loading amount
25 µg
Specification
fibroblasts, mitochondrial fraction
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Sep 29 2017

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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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

投稿 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:
Read More

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

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