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PhosphoTracer ERK1/2 pThr202/Tyr204 + Total ELISA Kit (ab119659)

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Overview

  • Product namePhosphoTracer ERK1/2 pThr202/Tyr204 + 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, Hamster, 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 ERK 1/2 assay kits detect endogenous levels of ERK 1/2 (GenBank Accessions NP_002737.2 [ERK1], NP_620407 [ERK2]) in cellular lysates. The phospho-ERK 1/2 assay detects ERK1 only when phosphorylated at Thr202/Tyr204, and ERK2 only when phosphorylated at Thr185/Tyr187. The total ERK assay detects ERK1/2 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.

  • Notes

    Sensitivity: Phospho-ERK 1/2: 10 pg/ml (Tested with rhERK1), ERK 1/2: 100 pg/ml (Tested with rhERK1).

    Range: Phospho-ERK 1/2: 10-10,000 pg/ml (Tested with rhERK1), ERK 1/2: 100-100,000 pg/ml (Tested with rhERK1).

     

  • Tested applicationsSandwich ELISA more details

Properties

  • FunctionInvolved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK-1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4).
  • Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • DomainThe TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications    Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-204.

    • Target information above from: UniProt accession P27361 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        ERKERK 2ERK-1
        ERK1ERK2ERT1ERT2Extracellular signal regulated kinase 2Extracellular signal-regulated kinase 1HS44KDAPHUMKER1AInsulin-stimulated MAP2 kinaseMAP kinase 1MAP kinase 2MAP kinase 3MAP kinase isoform p42MAP kinase isoform p44MAPK 1MAPK 3MAPK2Mapk3Microtubule-associated protein 2 kinaseMitogen activated protein kinase 2Mitogen-activated protein kinase 1Mitogen-activated protein kinase 3MK03_HUMANp38p40p41p41mapkP42MAPKp44-ERK1p44-MAPKP44ERK1P44MAPKPRKM1PRKM2PRKM3Protein tyrosine kinase ERK2
      see all
  • Database links
    see all

    • Applications

    Our Abpromise guarantee covers the use of ab119659 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-ERK 1/2: Tested in HeLa, A431, CHO; ERK 1/2: Tested in HEK293, HeLa, A431, C2C12, Raw264.7, A549, PC3, NIH3T3, U2OS.

    PhosphoTracer ERK1/2 pThr202/Tyr204 + Total ELISA Kit images

    • Sandwich ELISA - PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659)
      Sandwich ELISA - PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659)
      PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659) used in Sandwich ELISA.
    • Sandwich ELISA - PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659)
      Sandwich ELISA - PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659)
      Using the ERK 1/2 assay kits, a significant reduction of ERK 1/2 phosphorylation is detected in A431 cells treated with AG1478 for 3 hours compared with untreated cells, while only a small change in total ERK levels is observed.
    • Western blot - PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659)
      Western blot - PhosphoTracer ERK 1/2 (pT202/Y204) + total ERK ELISA Kit (ab119659)
      Using Western blot, a significant reduction of ERK 1/2 phosphorylation is detected in A431 cells treated with AG1478 for 3 hours compared with untreated cells, while only a small change in total ERK levels is observed.

    Protocols

    References for PhosphoTracer ERK1/2 pThr202/Tyr204 + Total ELISA Kit (ab119659)

    ab119659 has not yet been referenced specifically in any publications.

    Publishing research using ab119659 ? Please let us know so that we can cite the reference in this datasheet

    Product Wall

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    Displaying 1 - 7 of 7 results for Abreviews and Q&A

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    Votes: Highest First

    Customer kindly contacted us wondering why there is 3 times as much phospho specific antibody in the kit compared to the "total" antibody. Also, is the "total" antibody pair available for separate purchase so they can have the same amo...

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    The phospho/total combo kits are sold in a 3:1 ratio as we have found that users typically want more experiments for detecting phosphorylation than for detecting total ERK 1/2.

    While they do not have the separate total antibody pair available for...

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    What is the epitope of the capture and detection antibody?

    In the p-ERK assay, the capture antibody is specific for the phospho-T202/Y204 epitope, whereas the “total ERK” antibody is raised against a sequence near the C-terminus. I am afraid that we cannot be more precise than this, as the antibody is supplied fr...

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    Enhancer solution:
    1) function?
    2) composition?
    3) can be used with additional media but not in other cases - seems inconsistent/confusing



    1. Enhancer is to improve the sensitivity of most of the assays. It also plays a role in lysis, which is why it is added to the lysis buffer.

    2. We do not routinely release the composition of the buffers.

    3. The enhancer is cer...

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    1) I need to keep my cell sample before using the kit. How can I keep my cell sample (frozen at -20C)?

    2) How should I use the Assay Control Lysate? Should it be made in dilutions for comparison with my sample? Is my sample going to have a high...

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    Thank you for your enquiry.





    1) You can store the cells at -20C or -80C.

    2) The Assay Control Lysate is provided as a concentrated positive control to verify the ELISA kit works as intended. Once the lyophilised m...

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    Inquiry: What are the filter wavelength to read the fluoresence?

    Thank you for your enquiry.

    I can confirm this information is provided in the ADHP HRP substrate information section of the protocol provided under the protocols tab on the online datasheet:

    'Use with excitation filters in the range ...

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    Kits phosphotracer
    cibles ERK 1/2, p38 MAPKalpha, JNK 1/2/3.

    Merci de nous avoir contactés.

    Voici la liste des kits

    1) pour la détection d'une seule formed'une protéine particulière :

    ERK 1/2 (pThr202/Tyr204) : ab119625, http://www.abcam.com/ab119625
    ER...

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    Thanks for the information. I am interested in pERK ELISA kit as well as dual pAKT/pERK ELISA kit. Do you have them? Thank you,

    Thank you for your inquiry. I can recommend the following products: For pERK: ab119625 (PhosphoTracer ERK 1/2 (pT202/Y204) ELISA Kit) Click here (or use the following: http://www.abcam.com/index.html?datasheet=119625). ab119659 (PhosphoTracer ERK 1/2 (pT2...

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