製品の概要

  • 製品名
    Anti-Acetyl CoA synthetase antibody
    Acetyl CoA synthetase 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Acetyl CoA synthetase
  • アプリケーション
    適用あり: WB, ICC/IF, IHC-P, IPmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 650 to the C-terminus of Human Acetyl CoA synthetase.

    (Peptide available as ab71894.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in Human Liver and Human Colon Tissue lysates and in the following whole cell lysates: HepG2; U-87 MG; Caco 2.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab66038 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 79 kDa (predicted molecular weight: 79 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

ターゲット情報

  • 機能
    Activates acetate so that it can be used for lipid synthesis or for energy generation.
  • 配列類似性
    Belongs to the ATP-dependent AMP-binding enzyme family.
  • 細胞内局在
    Cytoplasm.
  • Information by UniProt
  • 参照データベース
  • 別名
    • ACAS2 antibody
    • AceCS antibody
    • Acetate CoA ligase antibody
    • Acetate thiokinase antibody
    • Acetate--CoA ligase antibody
    • Acetyl CoA synthetase antibody
    • Acetyl Coenzyme A synthetase 2 (ADP forming) antibody
    • Acetyl coenzyme A synthetase cytoplasmic antibody
    • Acetyl-CoA synthetase antibody
    • Acetyl-coenzyme A synthetase antibody
    • ACS antibody
    • ACSA antibody
    • ACSA_HUMAN antibody
    • ACSS2 antibody
    • Acyl activating enzyme antibody
    • Acyl CoA synthetase short chain family member 2 antibody
    • Acyl-activating enzyme antibody
    • Acyl-CoA synthetase short-chain family member 2 antibody
    • Cytoplasmic acetyl coenzyme A synthetase antibody
    • cytoplasmic antibody
    • MYH7B antibody
    see all

画像

  • All lanes : Anti-Acetyl CoA synthetase antibody (ab66038) 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
    Lane 3 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
    Lane 4 : Human colon tissue lysate - total protein (ab30051)
    Lane 5 : Caco 2 whole cell lysate (ab3950)

    Lysates/proteins at 10 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 79 kDa
    Observed band size : 79 kDa
    Additional bands at : 100 kDa,38 kDa,45 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • Acetyl CoA synthetase was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to Acetyl CoA synthetase and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab66038.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 79kDa: Acetyl CoA synthetase.
  • IHC image of Acetyl CoA synthetase staining in human liver carcinoma 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 ab66038, 5µg/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.
  • ICC/IF image of ab66038 stained HeLa cells. The cells were 4% PFA 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 (ab66038, 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) HeLa cells at 1µg/ml.

参考文献

This product has been referenced in:
  • Sun L  et al. Decreased expression of acetyl-CoA synthase 2 promotes metastasis and predicts poor prognosis in hepatocellular carcinoma. Cancer Sci 108:1338-1346 (2017). WB, IHC . Read more (PubMed: 28387999) »
  • Chen R  et al. The acetate/ACSS2 switch regulates HIF-2 stress signaling in the tumor cell microenvironment. PLoS One 10:e0116515 (2015). WB . Read more (PubMed: 25689462) »

See all 3 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Immuno-precipitation step
Other - Trueblot from rockland
Sample
Human Cell lysate - whole cell (human fibroblast)
Specification
human fibroblast
Total protein in input
500 µg
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Verified customer

投稿 Aug 20 2013

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (human fibroblast)
Specification
human fibroblast
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jun 10 2013

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

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