c-Jun pSer73 + Total PhosphoTracer ELISA Kit (ab119657)

Overview

  • Product namec-Jun pSer73 + Total PhosphoTracer ELISA Kit
  • Detection methodFluorescent
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSemi-quantitative
  • 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 c-Jun (p-Ser73) assay kits detect endogenous levels of c-Jun (GenBank Accession NP_002219) in cellular lysates. Phospho-c-Jun (p-Ser73) assay kits only detect c-Jun when phosphorylated at Ser73. Total c-Jun assay kits detect c-Jun 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-c-Jun: 3,000 cells/well (Tested in NIH3T3 cells), c-Jun: 3,000 cells/well (Tested in NIH3T3 cells).

    Range: Phospho-c-Jun: 3,000-100,000 cells/well tested (Tested in NIH3T3 cells), c-Jun: 3,000-100,000 cells/well tested (Tested in NIH3T3, cells).

  • Tested applicationsSandwich ELISAmore details
  • PlatformMicroplate

Properties

  • FunctionTranscription factor that recognizes and binds to the enhancer heptamer motif 5'-TGA[CG]TCA-3'.
  • Sequence similaritiesBelongs to the bZIP family. Jun subfamily.
    Contains 1 bZIP domain.
  • Post-translational
    modifications
    Phosphorylation enhances the transcriptional activity. Phosphorylated by PRKDC.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Alternative names
    • Activator protein 1
    • AP 1
    • AP1
    • Enhancer binding protein AP1
    • JUN
    • Jun A
    • Jun activation domain binding protein
    • Jun oncogene
    • Jun proto oncogene
    • JUN_HUMAN
    • Junc
    • Oncogene Jun
    • p39
    • Proto-oncogene c-jun
    • Transcription factor AP-1
    • v jun sarcoma virus 17 oncogene homolog
    • V-jun avian sarcoma virus 17 oncogene homolog
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab119657 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. Phospho-c-Jun (S73): Tested in HeLa, PC3, NIH3T3, HEK293; c-Jun: Tested in HeLa, PC3, NIH3T3, U2OS, HEK293.

c-Jun pSer73 + Total PhosphoTracer ELISA Kit images

  • Using Western blot, c-Jun phosphorylation at Ser73 is detected in anisomycin-treated NIH3T3 cells (+), compared with untreated cells (-), whereas no change in total c-Jun levels was observed.
  • Using the c-Jun (p-Ser73) assay kits, c-Jun phosphorylation at Ser73 is detected in anisomycin-treated NIH3T3 cells (+), compared with untreated cells (-), whereas no change in total c-JUN levels was observed.
  • Using the PhosphoTracer total-c-Jun assay, the expression of c-Jun was examined in several cell lines. Cells were seeded at 40K/well in 96-well microplates, and cultured overnight at 37°C in medium containing 10% FBS. The following day, the medium was removed from the wells, and the cells were lysed with 120 µL/well Lysis Mix, with shaking for 10 min. 50 µL of lysate was transferred to replicate wells of a PhosphoTracer assay plate, and assayed for total c-Jun using the standard PhosphoTracer protocol. Signal in the wells was determined using a plate reader. c-Jun was readily detected in most cell lines, with the exception of MCF7 and A431 cells.
  • HeLa cells were seeded at 40K cells/well in medium containing 10% FBS in a 96 well microplate, and cultured overnight. The following day the culture medium was removed, and the cells were treated with various concentrations of anisomycin diluted in serum-free medium for 30 mins. The medium was removed from the wells, and the cells were lysed with 120 µL/well Lysis Mix, with shaking for 10 min. 50 µL of lysate was transferred to replicate wells of a PhosphoTracer assay plate, and assayed for total c-Jun using the standard PhosphoTracer protocol. Signal in the wells was determined using a plate reader.

Protocols

References for c-Jun pSer73 + Total PhosphoTracer ELISA Kit (ab119657)

ab119657 has not yet been referenced specifically in any publications.

Product Wall

Thank you for your inquiry.

Unfortunately, the only ELISA kits we have to detect phospho c-jun in our catalog are ab119621, ab119622 and ab119657 which are not validated for blood plasma samples.

I have contacted the lab who confir...

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