Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) 抗体 [EPR933Y] (ab68476)

製品の概要

  • 製品名Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y]
    GSK3 (alpha + beta) 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR933Y] to GSK3 (alpha + beta) (phospho Y216 + Y279)
  • 特異性This antibody detects both GSK alpha phosphorylated on tyrosine 279 and GSK3 beta phosphorylyated on tyrosine 216.
  • アプリケーション適用あり: WB, IP, IHC-P, ICCmore details
    適用なし: Flow Cyt
  • 種交差性
    交差種: Mouse, Rat, Human, Zebrafish
  • 免疫原

    A synthetic phospho-peptide corresponding to residues surrounding tyrosine 279 in human GSK3 alpha and tyrosine 216 in human GSK beta.

  • ポジティブ・コントロール
    • WB: HEK-293 cell lysates, cell lysate from untreated and nerve growth factor-beta treated PC12 cells. IHC:human brain tissue.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab68476 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 1/500 - 1/2000. Detects a band of approximately 47-52 kDa (predicted molecular weight: 47-52 kDa).
IP 1/40.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC 1/250 - 1/500.
  • 追加情報Is unsuitable for Flow Cyt.
  • ターゲット情報

    • 関連性Glycogen synthase kinase 3 (GSK3) is a proline directed serine threonine kinase that was initially identified as a phosphorylating and inactivating glycogen synthase, a key enzyme in glycogen metabolism. Since then, it has been shown to be involved in the regulation of a diverse array of cellular functions, including protein synthesis, cell proliferation, cell differentiation, microtubule assembly/disassembly, and apoptosis. GSK3s substrate specificity is unique in that phosphorylation of substrate only occurs if a phosphoserine or phosphotyrosine is present four residues C terminal to the site of GSK phosphorylation. There exists two isoforms of GSK3, alpha and beta, and they show a high degree of amino acid homology. The two isoforms of GSK3 are strictly regulated via phosphorylation. Phosphorylation of GSK3 beta on Ser9 (Ser21 in GSK3 alpha) by protein kinase B (PKB) causes its inactivation is the primary mechanism responsible for growth factor inhibition of this kinase. Activation of GSK3 beta is dependent upon the phosphorylation of Tyr216 (Tyr279 in GSK3 alpha). Upon activation, it has been shown to phosphorylate a number of different cellular proteins, including p53, c-Myc, c-Jun, heat shock factor 1 (HSF1), and cyclin D1. GSK3 beta also has been shown to phosphorylate aberrant sites on the microtubule associated protein tau, which is critical for the progression of Alzheimer's disease. GSK3B is involved in energy metabolism, neuronal cell development, and body pattern formation.
    • 細胞内局在Cytoplasmic and Nuclear
    • 参照データベース
    • 別名
      • Factor A antibody
      • Glycogen synthase kinase 3 alpha antibody
      • Glycogen synthase kinase 3 beta antibody
      • GSK3 alpha antibody
      • GSK3 beta antibody
      • GSK3B antibody
      see all

    Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] 画像

    • All lanes : Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] (ab68476) at 1/500 dilution

      Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
      Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) treated with nerve growth factor at 100ng/ml for 5 minutes. Whole cell lysates.
      Lane 3 : SH-SY5Y (Human neuroblastoma epithelial cell) treated with nerve growth factor at 100ng/ml for 5 minutes. Whole cell lysates. Then the membrane was incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 47-52 kDa
      Observed band size : 47-52 kDa


      Exposure time : 10 seconds

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    • Human brain tissue stained with ab68476 at 1/100 dilution.
    • GSK3 (alpha + beta) (phospho Y216 + Y279) was immunoprecipitated from 10μg HeLa (human cervix adenocarcinoma) whole cell lysate with ab68476 at 1/50 dilution (2μg in 0.35mg lysates). Western blot was performed from the immunoprecipitate using ab68476 at 1/200 dilution (9 μg/ml). VeriBlot for IP (HRP) (ab131366), was used as the secondary antibody at 1/1000 dilution.

      Lane 1 (Input): HeLa (human cervix adenocarcinoma) treated with 1uM staurosporine for 4h whole cell lysate 10μg
      Lane 2 (+): HeLa (human cervix adenocarcinoma) treated with 1uM staurosporine for 4h whole cell lysate
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab68476 in HeLa (human cervix adenocarcinoma) treated with 1uM staurosporine for 4h whole cell lysate

      Exposure Time: 30 seconds
      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

       

    • All lanes : Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] (ab68476)

      Lane 1 : PC12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 2 : Whole cell lysate from PC12 (Rat adrenal gland pheochromocytoma cell line) cells treated with nerve growth factor-ß at 100ng/ml for 5 minutes.
      Lane 3 : Whole cell lysate from PC12 (Rat adrenal gland pheochromocytoma cell line) cells ) treated with nerve growth factor-ß at 100ng/ml for 5 minutes. Membrane incubated with phosphatase

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG H+L (HRP))
      Developed using the ECL technique

      Predicted band size : 47-52 kDa


      Exposure time : 3 seconds

      Blocking and dilution buffer: 5% NFDM/TBST

    • Dot Blot analysis of Lane 1: GSK3 (alpha + beta) (pY216 + pY279) phospho peptide and Lane 2: GSK3 (alpha + beta) non-phospho peptide labeling GSK3 (alpha + beta) (phospho Y216 + Y279) with ab68476 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.

    • All lanes : Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] (ab68476) at 1/2000 dilution

      Lane 1 : 293 cell lysate, cells untreated.
      Lane 2 : 293 cell lysate, cells treated with AP.

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP conjugated Goat anti-rabbit at 1/2000 dilution

      Predicted band size : 47-52 kDa
      Observed band size : 47-52 kDa

    Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] (ab68476) 使用論文

    This product has been referenced in:
    • Xu R  et al. The protease Omi regulates mitochondrial biogenesis through the GSK3ß/PGC-1a pathway. Cell Death Dis 5:e1373 (2014). WB ; Mouse . Read more (PubMed: 25118933) »
    • Weigand S  et al. Global quantitative phosphoproteome analysis of human tumor xenografts treated with a CD44 antagonist. Cancer Res 72:4329-39 (2012). Read more (PubMed: 22777824) »

    See all 4 Publications for this product

    Product Wall

    Application Immunohistochemistry free floating
    Sample Mouse Tissue sections (Brain)
    Specification Brain
    Username

    Abcam user community

    Verified customer

    投稿 Jan 14 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry free floating
    Sample Mouse Tissue sections (Hippocampus CA1 mouse Brain)
    Specification Hippocampus CA1 mouse Brain
    Username

    Abcam user community

    Verified customer

    投稿 Jan 19 2016

    Application Immunohistochemistry (Frozen sections)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Sample Zebrafish Tissue sections (retina)
    Specification retina
    Permeabilization Yes - triton X
    Fixative Paraformaldehyde
    Username

    Dr. Ryan MacDonald

    Verified customer

    投稿 May 22 2014

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