製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to ATP5A
  • アプリケーション
    適用あり: WB, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Chicken, Cow, Chimpanzee, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human ATP5A.

  • ポジティブ・コントロール
    • WB: Human fetal brain and human fetal heart tissue lysates and Raw264.7, HEK293, MCF7, HepG2, HL60 and PC12 whole cell lysates. IHC-P: Human heart muscle tissue sections. ICC/IF: HeLa cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab151229 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 59 kDa (predicted molecular weight: 59 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.

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

画像

  • ICC/IF image of ab151229 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab151229, 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.

    This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% formaldehyde fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.

  • All lanes : Anti-ATP5A antibody (ab151229) at 1 µg/ml

    Lane 1 : Brain (Human) Tissue Lysate - fetal normal tissue (ab29467)
    Lane 2 : Heart (Human) Whole Cell Lysate - fetal normal tissue
    Lane 3 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 7 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate
    Lane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 59 kDa
    Observed band size : 59 kDa
    Additional bands at : 37 kDa,50 kDa,75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 secondsThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab151229 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
  • IHC image of ATP5A staining in human heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab151229, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

参考文献

ab151229 has not yet been referenced specifically in any publications.

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