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PhosphoTracer βCatenin pSer45 + Total ELISA Kit (ab119656)

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Overview

  • Product namePhosphoTracer βCatenin pSer45 + Total ELISA Kit
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSandwich
  • Assay time
    1h 15m
  • 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 assay kits detect endogenous levels of beta-catenin (GenBank Accession NP_001895.1) in cellular lysates. Phospho-beta-catenin assays only detect beta-catenin when phosphorylated at Ser45. Total beta-catenin assays detect beta-catenin levels irrespective of phosphorylation.



    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.

  • Notes

    Sensitivity: Phospho-AKT: 10,000 cells/well (Tested in A549 cells), AKT: 2,000 cells/well (Tested in A549 cells).

    Range: Phospho-AKT: 10,000-100,000 cells/well tested (Tested in A549 cells), AKT: 2,000-100,000 cells/well tested (Tested in A549 cells).

  • Tested applicationsSandwich ELISA more details

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.

    • Target information above from: UniProt accession P35222 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        Beta-cateninBfcCadherin associated protein beta
        Catenin (cadherin associated protein) beta 1Catenin beta-1CatnbCTNB1_HUMANCTNNBCTNNB1Mesc
      see all
  • Database links

      • Applications

      Our Abpromise guarantee covers the use of ab119656 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 sELISA. Phospho-beta-Catenin (S45): Tested in A549, C2C12; beta-Catenin: Tested in HEK293, HeLa, MCF7, A431, A549, PC3, NIH3T3, C2C12

      PhosphoTracer βCatenin pSer45 + Total ELISA Kit images

      • Western blot - PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656)
        Western blot - PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656)
        Using Western blot, beta-catenin phosphorylation at Ser45 is detected in PMA/calyculin-treated A549 cells (+), compared with untreated A549 cells (-).
      • Sandwich ELISA - PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656)
        Sandwich ELISA - PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656)
        Using the beta-catenin assay kits, beta-catenin phosphorylation at Ser45 is detected in PMA/calyculin-treated A549 cells (+), compared with untreated A549 cells (-).
      • Sandwich ELISA - PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656)
        Sandwich ELISA - PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656)
        PhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA Kit (ab119656) used in Sandwich ELISA.

      Protocols

      References for PhosphoTracer βCatenin pSer45 + Total ELISA Kit (ab119656)

      ab119656 has not yet been referenced specifically in any publications.

      Publishing research using ab119656 ? Please let us know so that we can cite the reference in this datasheet

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      Llamada de telefono.
      Se miden valores absolutos? al no haber una recta de estandares para comparar...
      Pone "leer a la longitud de onda adecuada": Cual es? la de emision? emision menos absorcion...?

      Gracias por contactar con Abcam.

      He recibido una respuesta de mis compañeros del laboratorio. Me indican que se miden valores absolutos, es un ensayo semi cuantitativo, y dichos valores se usan como una estimación en el incremento ...

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      What do you mean with ab119620 detects total beta catenin, could you please specify your answer? Does this kit detect both the active (non-p) form and the phosphorylated form, in that case there is no difference between the ab119620 and ab119656? Or does ...

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      Thank you for your reply.

      ab119620 will detect total phospho beta-catenin regardless of phosphorylation state. It will detect both non-phosphorylated and phosphorylated beta-catenin in the same well. 1 antibody is included in the kit.
      Read More

      I've some questions regarding the following products:PhosphoTracer beta-Catenin Total ELISA kit (ab119620) andPhosphoTracer beta-Catenin (pS45) + total beta-Catenin ELISA kit(ab119656).


      - What is the differences between these two kits?

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      Thank you for your email.

      I have answered your questions below:

      - What is the differences between these two kits?
      ab119620 detects total beta catenin
      ab119656 detects both total beta catenin and phosphorylated beta catenin...

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