製品の概要

  • 製品名Anti-MGMT antibody
    MGMT 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to MGMT
  • アプリケーション適用あり: WB, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human MGMT.

    (Peptide available as ab89380.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Irradiated HeLa; Jurkat; HepG2; Caco2. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal tonsil. ICC/IF: HAP1 wild-type and HAP1-MGMT knockout cells

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab80513 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 25 kDa (predicted molecular weight: 21 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ターゲット情報

  • 機能Involved in the cellular defense against the biological effects of O6-methylguanine (O6-MeG) in DNA. Repairs alkylated guanine in DNA by stoichiometrically transferring the alkyl group at the O-6 position to a cysteine residue in the enzyme. This is a suicide reaction: the enzyme is irreversibly inactivated.
  • 配列類似性Belongs to the MGMT family.
  • 細胞内局在Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 6 O methylguanine DNA methyltransferase antibody
    • 6-O-methylguanine-DNA methyltransferase antibody
    • Agat antibody
    • AGT antibody
    • AI267024 antibody
    • EC 2.1.1.63 antibody
    • Methylated DNA protein cysteine methyltransferase antibody
    • Methylated-DNA--protein-cysteine methyltransferase antibody
    • Methylguanine DNA methyltransferase antibody
    • MGC107020 antibody
    • MGMT antibody
    • MGMT_HUMAN antibody
    • O 6 methylguanine DNA alkyltransferase antibody
    • O 6 methylguanine DNA methyltransferase antibody
    • O-6-methylguanine-DNA methyltransferase antibody
    • O-6-methylguanine-DNA-alkyltransferase antibody
    see all

Anti-MGMT antibody 画像



  • Predicted band size : 21 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: MGMT knockout HAP1 cell lysate (20 µg)

    Lane 3: MCF7 cell lysate (20 µg)
    Lane 4: Jurkat cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab80513 observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab80513 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab80513 and ab8245 (loading control to GAPDH) were diluted 1μg/mL and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab80513 staining MGMT in wild-type HAP1 cells (top panel) and MGMT knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), 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 with ab80513 at 1μg/ml concentration and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • IHC image of MGMT staining in Human normal tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab80513, 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.

     

    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.

  • All lanes : Anti-MGMT antibody (ab80513) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Irradiated HeLa Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 5 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 21 kDa
    Observed band size : 25 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 120 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes
    Methylated-DNA--protein-cysteine methyltransferase (MGMT) contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

Anti-MGMT antibody (ab80513) 使用論文

ab80513 has not yet been referenced specifically in any publications.

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