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Overview

  • Product namePhosphoTracer GAPDH ELISA Kit
  • Detection methodFluorescent
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSandwich
  • Sensitivity
    25 pg/ml
  • Range
    25 pg/ml - 6000 pg/ml
  • Assay time
    1h 15m
  • 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 GAPDH assay kits detect endogenous levels of GAPDH (GenBank Accession NP_002037) in cellular lysates.



    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

  • 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 GAPDH (HRP) (96 assay points) 1 x 3ml
    Rabbit monoclonal GAPDH (96 assay points) 1 x 3ml
    Stop Solution 1 x 2ml
    Wash Buffer (10X) 1 x 15ml
  • Research Areas
  • FunctionHas both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.
  • PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
  • Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
  • Post-translational
    modifications    S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
    ISGylated.
  • Cellular localizationCytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.

    • Target information above from: UniProt accession P04406 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        Aging associated gene 9 proteinG3P_HUMANG3PD
        GAPDGAPDHGlyceraldehyde 3 phosphate dehydrogenaseGlyceraldehyde-3-phosphate dehydrogenasePeptidyl-cysteine S-nitrosylase GAPDH
      see all
  • Database links

      • Applications

      Our Abpromise guarantee covers the use of ab119627 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 in HeLa, A431, A549, PC3, U2OS.

      PhosphoTracer GAPDH ELISA Kit images

      • Sandwich ELISA - PhosphoTracer GAPDH ELISA kit (ab119627)
        Sandwich ELISA - PhosphoTracer GAPDH ELISA kit (ab119627)
        As shown, using the GAPDH assay kit, cellular GAPDH is readily detected in A431 cellular lysates, in either untreated cells (-), or cells treated with EGF (+).
      • Sandwich ELISA - PhosphoTracer GAPDH ELISA kit (ab119627)
        Sandwich ELISA - PhosphoTracer GAPDH ELISA kit (ab119627)
        Various concentrations of recombinant GAPDH diluted in Lysis Mix containing 0.1% BSA were analyzed using the PhosphoTracer cellular GAPDH assay. The recombinant GAPDH dilutions were transfered to an PhosphoTracer assay plate and analyzed using the standard PhosphoTracer assay protocol. Signal in the wells was determined using a plate reader.
      • Sandwich ELISA - PhosphoTracer GAPDH ELISA kit (ab119627)
        Sandwich ELISA - PhosphoTracer GAPDH ELISA kit (ab119627)
        Using the PhosphoTracer GAPDH assay, the amount of lysate required to detect GAPDH in HeLa cell lysates was examined. Up to 50 µg of lysate/well was loaded into replicate wells of an PhosphoTracer microplate, and analyzed for GAPDH. GAPDH was readily detected in less than 1 µg of lysate/well.

      Protocols

      References for PhosphoTracer GAPDH ELISA Kit (ab119627)

      ab119627 has not yet been referenced specifically in any publications.

      Publishing research using ab119627 ? Please let us know so that we can cite the reference in this datasheet

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      Votes: Highest First

      J’ai acheté le kit « PhosphoTracer GAPDH ELISA kit » (réf : ab119627)
      Pour mes applications, j’ai besoin de faire une gamme de GAPDH.
      Pouvez-vous me donner une référence de standard GAPDH (humai...

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      Merci de nous avoir contactés.

      Je recommanderais la GAPDH référence catalogue ab82633. Le lien vers la fiche technique est http://www.abcam.com/ab82633.

      J'espère que ces informations vous seront utiles. N'h&ea...

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      Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"