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Overview

  • Product namePhosphoTracer BAD Total ELISA Kit
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSandwich
  • Sensitivity
    2500 cells/well
  • Range
    2500 cells/well - 80000 cells/well
  • 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 PhospoTracer BAD Total assay kits are for detection of endogenous and transfected levels of BAD (GenBank Accession NP_004313) in cellular lysates. Total BAD assay kits detect BAD irrespective of phosphorylation status.



    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

Properties

  • Storage instructionsStore at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    96-well PhosphoTracer assay plate (stripwell) 1 unit
    Adherent plate seal 2 units
    ADHP (100X) 1 x 120µl
    ADHP Dilution Buffer 1 x 15ml
    Assay Control Lysate (lyophilized) 1 x 0.25ml
    Enhancer Solution 1 x 1ml
    Lysis Buffer (5X) 1 x 15ml
    Mouse monoclonal BAD (HRP) (96 assay points) 1 x 3ml
    Rabbit monoclonal BAD (96 assay points) 1 x 3ml
    Stop Solution 1 x 2ml
    Wash Buffer (10X) 1 x 15ml
  • Research Areas
  • FunctionPromotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2 (By similarity). Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.
  • Tissue specificityExpressed in a wide variety of tissues.
  • Sequence similaritiesBelongs to the Bcl-2 family.
  • DomainIntact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family.
  • Post-translational
    modifications    Phosphorylated on one or more of Ser-75, Ser-99, Ser-118 and Ser-134 in response to survival stimuli, which blocks its pro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75 promotes heterodimerization with 14-3-3 proteins. This interaction then facilitates the phosphorylation at Ser-118, a site within the BH3 motif, leading to the release of Bcl-X(L) and the promotion of cell survival. Ser-99 is the major site of AKT/PKB phosphorylation, Ser-118 the major site of protein kinase A (CAPK) phosphorylation. Ser-75 is phosphorylated by AKT/PKB, protein kinase A and PIM2.
  • Cellular localizationMitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm.

    • Target information above from: UniProt accession Q92934 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        BADBAD_HUMANBBC2
        BBC6BCL X/BCL 2 binding proteinBcl-2-binding component 6Bcl-2-like protein 8Bcl-XL/Bcl-2-associated death promoterBcl2 antagonist of cell deathBCL2 antagonist of cell death proteinBCL2 associated agonist of cell deathBCL2 binding component 6BCL2 binding proteinBcl2-L-8BCL2L8
      see all
  • Database links

      • Applications

      Our Abpromise guarantee covers the use of ab119618 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. Tested on PC3, MCF7, HEK293, HeLa, NIH3T3

      PhosphoTracer BAD Total ELISA Kit images

      • Sandwich ELISA - PhosphoTracer BAD Total ELISA kit (ab119618)
        Sandwich ELISA - PhosphoTracer BAD Total ELISA kit (ab119618)
        Detection of total BAD in PC3 cells treated with various concentrations of UCN-01, using ab119618.
      • Sandwich ELISA - PhosphoTracer BAD Total ELISA kit (ab119618)
        Sandwich ELISA - PhosphoTracer BAD Total ELISA kit (ab119618)
        Detection of total BAD in several different cell lines, using ab119618.

      Protocols

      References for PhosphoTracer BAD Total ELISA Kit (ab119618)

      ab119618 has not yet been referenced specifically in any publications.

      Publishing research using ab119618 ? Please let us know so that we can cite the reference in this datasheet

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