beta-Catenin (pS45) PhosphoTracer ELISA Kit (ab119619)

Overview

  • Product namebeta-Catenin (pS45) PhosphoTracer ELISA Kit
  • Detection methodFluorescent
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSemi-quantitative
  • Sensitivity
    10000 cells/well
  • Range
    10000 cells/well - 100000 cells/well
  • 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 beta-catenin (pS45) assay kits detect endogenous levels of beta-catenin (GenBank Accession NP_001895.1) in cellular lysates. Phospho-beta-catenin assay kits only detect beta-catenin when phosphorylated at Ser45.



    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 ELISAmore details
  • PlatformMicroplate

Properties

  • FunctionKey dowstream component of the canonical Wnt signaling pathway. In the absence of Wnt, forms a complex with AXIN1, AXIN2, APC, CSNK1A1 and GSK3B that promotes phosphorylation on N-terminal Ser and Thr residues and ubiquitination of CTNNB1 via BTRC and its subsequent degradation by the proteasome. In the presence of Wnt ligand, CTNNB1 is not ubiquitinated and accumulates in the nucleus, where it acts as a coactivator for transcription factors of the TCF/LEF family, leading to activate Wnt responsive genes.
    Involved in the regulation of cell adhesion. The majority of beta-catenin is localized to the cell membrane and is part of E-cadherin/catenin adhesion complexes which are proposed to couple cadherins to the actin cytoskeleton.
  • Tissue specificityExpressed in several hair follicle cell types: basal and peripheral matrix cells, and cells of the outer and inner root sheaths. Expressed in colon.
  • Involvement in diseaseDefects in CTNNB1 are associated with colorectal cancer (CRC) [MIM:114500].
    Note=Activating mutations in CTNNB1 have oncogenic activity resulting in tumor development. Somatic mutations are found in various tumor types, including colon cancers, ovarian and prostate carcinomas, hepatoblastoma (HB), hepatocellular carcinoma (HCC). HBs are malignant embryonal tumors mainly affecting young children in the first three years of life.
    Defects in CTNNB1 are a cause of pilomatrixoma (PTR) [MIM:132600]; a common benign skin tumor.
    Defects in CTNNB1 are a cause of medulloblastoma (MDB) [MIM:155255]. MDB is a malignant, invasive embryonal tumor of the cerebellum with a preferential manifestation in children.
    Defects in CTNNB1 are a cause of susceptibility to ovarian cancer (OC) [MIM:167000]. Ovarian cancer common malignancy originating from ovarian tissue. Although many histologic types of ovarian neoplasms have been described, epithelial ovarian carcinoma is the most common form. Ovarian cancers are often asymptomatic and the recognized signs and symptoms, even of late-stage disease, are vague. Consequently, most patients are diagnosed with advanced disease.
    Note=A chromosomal aberration involving CTNNB1 is found in salivary gland pleiomorphic adenomas, the most common benign epithelial tumors of the salivary gland. Translocation t(3;8)(p21;q12) with PLAG1.
  • Sequence similaritiesBelongs to the beta-catenin family.
    Contains 12 ARM repeats.
  • Post-translational
    modifications
    Phosphorylation by GSK3B requires prior phosphorylation of Ser-45 by another kinase. Phosphorylation proceeds then from Thr-41 to Ser-37 and Ser-33.
    EGF stimulates tyrosine phosphorylation. Phosphorylation on Tyr-654 decreases CDH1 binding and enhances TBP binding.
    Ubiquitinated by the SCF(BTRC) E3 ligase complex when phosphorylated by GSK3B, leading to its degradation. Ubiquitinated by a E3 ubiquitin ligase complex containing UBE2D1, SIAH1, CACYBP/SIP, SKP1, APC and TBL1X, leading to its subsequent proteasomal degradation.
  • Cellular localizationCytoplasm. Nucleus. Cytoplasm > cytoskeleton. Cell junction > adherens junction. Cell junction. Cell membrane. Cytoplasmic when it is unstabilized (high level of phosphorylation) or bound to CDH1. Translocates to the nucleus when it is stabilized (low level of phosphorylation). Interaction with GLIS2 and MUC1 promotes nuclear translocation. Interaction with EMD inhibits nuclear localization.
  • Information by UniProt
  • Alternative names
    • Beta-catenin
    • Bfc
    • Cadherin-associated protein, beta
    • Catenin (cadherin associated protein) beta 1 88kD
    • Catenin (cadherin associated protein), beta 1
    • Catenin beta-1
    • Catnb
    • CTNB1_HUMAN
    • CTNNB
    • CTNNB1
    • Mesc
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab119619 in the following tested applications.

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

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration. Tested in A549, C2C12

beta-Catenin (pS45) PhosphoTracer ELISA Kit images

  • Using Western blot, beta-catenin phosphorylation at Ser45 is detected in PMA/calyculin-treated A549 cells (+), compared with untreated A549 cells (-).
  • Using the beta-catenin assay kits, beta-catenin phosphorylation at Ser45 is detected in PMA/calyculin-treated A549 cells (+), compared with untreated A549 cells (-), consistent with Western blot results.
  • PhosphoTracer beta-Catenin (pS45) ELISA Kit (ab119619) used in Sandwich ELISA.

Protocols

References for beta-Catenin (pS45) PhosphoTracer ELISA Kit (ab119619)

ab119619 has not yet been referenced specifically in any publications.

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