PhosphoTracer SMAD1 (pS463/465) + SMAD3 (pS423/425) ELISA Kit (ab119680)

Overview

• Product namePhosphoTracer SMAD1 (pS463/465) + SMAD3 (pS423/425) 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.

   

  SMAD assay kits are for detection of endogenous levels of SMAD1 (GenBank Accession NP_001003688) and SMAD3 (GenBank Accession NP_001138574) in cellular lysates. The phospho-SMAD1 assay detects SMAD1 only when phosphorylated at Ser463/465. Based on sequence similarity, cross reaction to p-SMAD5 and p-SMAD8 may occur. The phospho-SMAD3 assay detects SMAD3 only when phosphorylated at Ser423/425. Based on sequence similarity, cross reaction to p-SMAD2 may occur.
  
  
  
  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-SMAD1: 5,000 cells/well (tested in C2C12 cells), Phospho-SMAD3: 5,000 cells/well (tested in C2C12 cells).

  Range: Phospho-SMAD1: 5,000-100,000 cells/well (tested in C2C12 cells), Phospho-SMAD3: 5,000-100,000 cells/well (tested in C2C12 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 Phospho-Smad 1(HRP) (48 assay points)	1 x 1.5ml
  Mouse monoclonal Phospho-Smad 3 (HRP) (48 assay points)	1 x 1.5ml
  Rabbit monoclonal Phospho Smad 1 (Ser463/465) (48 assay points)	1 x 1.5ml
  Rabbit monoclonal Phospho Smad 3 (Ser423/425) (48 assay points)	1 x 1.5ml
  Stop Solution	1 x 2ml
  Wash Buffer (10X)	1 x 15ml

• Research Areas
  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Hypoxia

  • Stem Cells
  • Signaling Pathways
  • TGF beta
  • Cytoplasmic

  • Stem Cells
  • Signaling Pathways
  • TGF beta
  • Nuclear

  • Cancer
  • Signal transduction
  • Nuclear signaling
  • SMAD family

  • Cancer
  • Cancer Metabolism
  • Response to hypoxia

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

  • Signal Transduction
  • Signaling Pathway
  • Nuclear Signaling
  • SMADs

  • Signal Transduction
  • Growth Factors/Hormones
  • TGF

  • Epigenetics and Nuclear Signaling
  • Nuclear Signaling Pathways
  • SMADs

Applications

PhosphoTracer SMAD1 (pS463/465) + SMAD3 (pS423/425) ELISA Kit images

• 

  Sandwich ELISA - PhosphoTracer SMAD1 (pS463/465) + SMAD3 (pS423/425) ELISA Kit (ab119680)
  Using the SMAD1 assay kits or Western blot, SMAD1 phosphorylation at Ser463/465 is detected in BMP4-treated C2C12 cells (+), compared with C2C12 cells treated with 50 µM dorsomorphin (-).
• 

  Sandwich ELISA - PhosphoTracer SMAD1 (pS463/465) + SMAD3 (pS423/425) ELISA Kit (ab119680)
  Using the p-SMAD3 assay kit or Western blot, SMAD3 phosphorylation at Ser423/425 is detected in TGF-treated C2C12 cells (+), compared with untreated C2C12 cells (-).

Protocols

Protocol Booklet