Overview

  • Product nameGSK3 beta Total PhosphoTracer ELISA Kit
  • Detection methodFluorescent
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSemi-quantitative
  • Sensitivity
    0.15 ng/ml
  • Range
    0.15 ng/ml - 500 ng/ml
  • Assay time
    2h 0m
  • Assay durationOne step assay
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    PhosphoTracer assays use a traditional immuno-sandwich format, but with a major difference both the analyte and the assay reagents are added to the PhosphoTracer assay microplate at the same time. After a short incubation period, unbound assay reagents and analytes are washed away, and immuno-complexes containing both antibodies are detected. The process can take as little as 60 minutes to complete.

    PhosphoTracer kits also allow a higher degree of assay flexibility. In contrast to other ELISA formats, no antibodies are present on the assay microplate itself, so assays for several different targets can be performed in different wells on the same microplate. Simply mix the lysate with your target reagents of choice, using the microplate configuration of your choice.

    A whole new way of performing cellular assays, PhosphoTracer takes the hard work out of running a standard ELISA, while still giving the high quality results expected from a sandwich immunoassay. Fully self-contained kits are supplied in convenient 96-well packs. Simple to use and highly sensitive PhosphoTracer kits are designed to get results, fast.

     

    Abcam's PhosphoTracer GSK3beta assay kits detect endogenous levels of GSK3beta (GenBank Accession NP_001139628) in cellular lysates. Total GSK3beta assay kits detect GSK3beta irrespective of phosphorylated at Ser9.



    The substrate used with the HRP conjugated detection antibody is a combination of 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP) (wavelength exc/em = 530-540nm / 590-600nm), a highly sensitive and stable substrate for HRP) and ADHP Dilution Buffer (a stabilized H2O2 solution). Learn more about the fluorogenic substrate, ADHP.

  • Tested applicationsSandwich ELISA more details
  • PlatformMicroplate

Properties

  • FunctionParticipates in the Wnt signaling pathway. Implicated in the hormonal control of several regulatory proteins including glycogen synthase, MYB and the transcription factor JUN. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Phosphorylates MUC1 in breast cancer cells, and decreases the interaction of MUC1 with CTNNB1/beta-catenin. Phosphorylates CTNNB1/beta-catenin. Phosphorylates SNAI1. Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. Prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization. Phosphorylates MACF1 and this phosphorylation inhibits the binding of MACF1 to microtubules which is critical for its role in bulge stem cell migration and skin wound repair.
  • Tissue specificityExpressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney.
  • Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Phosphorylated by AKT1 and ILK1. Activated by phosphorylation at Tyr-216.
  • Cellular localizationCytoplasm. Nucleus. Cell membrane. The phosphorylated form shows localization to cytoplasm and cell membrane. The MEMO1-RHOA-DIAPH1 signaling pathway controls localization of the phosophorylated form to the cell membrane.
  • Target information above from: UniProt accession P49841 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names
    • Glycogen synthase kinase-3 beta
    • GSK-3 beta
    • GSK3B
    • GSK3B_HUMAN
    • Serine/threonine-protein kinase GSK3B
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab119629 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Sandwich ELISA Use at an assay dependent concentration. Tested in A549, MCF7, NIH3T3

GSK3 beta Total PhosphoTracer ELISA Kit images

  • Using the GSK3beta assay kits, GSK3beta phosphorylation at Ser9 is detected in insulin-treated MCF7 cells (+), compared with cells treated with UCN-01 (-), whereas no change in total GSK3beta levels was observed.
  • Detection of total GSK3 beta, using ab119629 on NIH3T3 cells treated with varying concentrations of UCN-01 for 2 hours, then stimulated with insulin.
  • Using Western blot, GSK3beta phosphorylation at Ser9 is detected in insulin-treated MCF7 cells (+), compared with cells treated with UCN-01 (-), whereas no change in total GSK3beta levels was observed.

Protocols

References for GSK3 beta Total PhosphoTracer ELISA Kit (ab119629)

ab119629 has not yet been referenced specifically in any publications.

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