Anti-Cofilin (phospho S3) 抗体 (ab100836)

製品の概要

  • 製品名Anti-Cofilin (phospho S3) antibody
    Cofilin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Cofilin (phospho S3)
  • アプリケーション適用あり: ICC/IF, ELISA, IHC-P, WBmore 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 Cofilin, phosphorylated at S3.

  • ポジティブ・コントロール
    • WB: Jurkat and SHSY5Y whole cell lysates. IHC-P: FFPE human breast adenocarcinoma tissue. ICC/IF: MCF7 cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab100836 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
ICC/IF Use a concentration of 5 µg/ml.
ELISA Use at an assay dependent concentration.
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.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 20 kDa (predicted molecular weight: 18 kDa).

ターゲット情報

  • 機能Controls reversibly actin polymerization and depolymerization in a pH-sensitive manner. It has the ability to bind G- and F-actin in a 1:1 ratio of cofilin to actin. It is the major component of intranuclear and cytoplasmic actin rods.
  • 組織特異性Widely distributed in various tissues.
  • 配列類似性Belongs to the actin-binding proteins ADF family.
    Contains 1 ADF-H domain.
  • 翻訳後修飾Phosphorylated on Ser-3 in resting cells.
  • 細胞内局在Nucleus matrix. Cytoplasm > cytoskeleton. Almost completely in nucleus in cells exposed to heat shock or 10% dimethyl sulfoxide.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 18 kDa phosphoprotein antibody
    • CFL 1 antibody
    • CFL antibody
    • CFL1 antibody
    • COF1_HUMAN antibody
    • Cofilin 1 antibody
    • Cofilin 1 non muscle antibody
    • Cofilin antibody
    • Cofilin non muscle isoform antibody
    • Cofilin-1 antibody
    • epididymis secretory protein Li 15 antibody
    • HEL-S-15 antibody
    • non-muscle isoform antibody
    • p18 antibody
    see all

Anti-Cofilin (phospho S3) antibody 画像

  • ICC/IF image of ab100836 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 (ab100836, 5µg/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 4% formaldehyde fixed (10 min) HeLa cells at 5µg/ml,.

  • All lanes : Anti-Cofilin (phospho S3) antibody (ab100836) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
    Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
    Lane 6 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 18 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 110 kDa,150 kDa,33 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 180 seconds
  • IHC image of ab100836 staining in human breast adenocarcinoma 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 ab100836, 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.
  • ab100836 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.

Anti-Cofilin (phospho S3) antibody (ab100836) 使用論文

ab100836 has not yet been referenced specifically in any publications.

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