リコンビナント
RabMAb

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-Pmore details
    適用なし: Flow Cyt or ICC
  • 種交差性
    交差種: Mouse, Rat, Human, Zebrafish
  • 免疫原

    corresponding to Human GSK3 (alpha + beta).
    (Peptide available as ab186139)

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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This antibody may not be suitable for IHC with mouse, rat or zebrafish samples.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/100 - 1/250.

  • 追加情報
    Is unsuitable for Flow Cyt or ICC.
  • 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

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labeling GSK3 (alpha + beta) with Purified ab68476 at 1:50 dilution (5.3 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • All lanes : Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] (ab68476) at 1/1000 dilution (purified)

      Lane 1 : Mouse brain lysates. Then the membrane was incubated with phosphatase
      Lane 2 : Untreated Mouse brain lysates

      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

      Blocking and diluting buffer: 5% NFDM/TBST

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

      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.

    • All lanes : Anti-GSK3 (alpha + beta) (phospho Y216 + Y279) antibody [EPR933Y] (ab68476) at 1/1000 dilution (purified)

      Lane 1 : Zebrafish lysates Then the membrane was incubated with phosphatase
      Lane 2 : Untreated Zebrafish lysates

      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

      Blocking and diluting buffer: 5% NFDM/TBST

       

    • Human brain tissue stained with unpurified 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) (unpurified)

      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 (unpurified)

      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

    参考文献

    This product has been referenced in:
    • Roettger A  et al. Inhibitory Effect of 1,8-Cineol on ß-Catenin Regulation, WNT11 Expression, and Cellular Progression in HNSCC. Front Oncol 7:92 (2017). Read more (PubMed: 28589081) »
    • 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) »

    See all 5 Publications for this product

    レビューと Q&A

    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"

    登録