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

Overview

• Product namePhosphoTracer c-Jun pSer73 + 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 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 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 c-Jun (HRP)(24 assay points)	1 x 0.75ml
  Mouse monoclonal Phospho-c-Jun (HRP) (72 assay points)	3 x 0.75ml
  Rabbit monoclonal c-Jun (24 assay points)	1 x 0.75ml
  Rabbit polyclonal Phospho-c-Jun (Ser73)(72 assay points)	3 x 0.75ml
  Stop Solution	1 x 2ml
  Wash Buffer (10X)	1 x 15ml

• Research Areas
  • Kits/ Lysates/ Other
  • Kits
  • ELISA Kits
  • ELISA Kits
  • Phosphoprotein ELISA kits

  • Epigenetics and Nuclear Signaling
  • Transcription
  • Transcription Factors

Applications

PhosphoTracer c-Jun pSer73 + Total ELISA Kit images

• 

  Western blot - PhosphoTracer c-Jun (pS73) + total c-Jun ELISA Kit (ab119657)
  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.
• 

  Sandwich ELISA - PhosphoTracer c-Jun (pS73) + total c-Jun ELISA Kit (ab119657)
  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.
• 

  Sandwich ELISA - PhosphoTracer c-Jun (pS73) + total c-Jun ELISA Kit (ab119657)
  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.
• 

  Sandwich ELISA - PhosphoTracer c-Jun (pS73) + total c-Jun ELISA kit (ab119657)
  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

Protocol Booklet