Introduction
DuoSet IC (IntraCellular) Assay Development Systems offer a fast, sensitive, and economical method to detect and quantify intracellular factors involved in apoptosis, signal transduction, and transcription. Extensive validation work is done in-house to ensure specificity and detailed protocols are provided. Kits are available in 2-, 5-, and 15-(96 well) plate pack sizes. Kit contents vary depending on the assay format.
For more information about our complete ELISA kits for signal transduction molecules please see our Surveyor IC ELISA kit products
DuoSet IC Advantages
- Optimized reagents save lengthy development time.
- High specificity
- Available in 2-, 5-, and 15-96 well plate pack sizes*
ELISAs
A sensitive, convenient, and high-throughput alternative to Western blots, these kits contain Capture Antibody, Standard, Biotinylated Detection Antibody, and Streptavidin-HRP. Target protein, either total or phosphorylated, is sandwiched between an immobilized Capture Antibody and a Biotinylated Detection Antibody. Signal is generated using Streptavidin HRP and substrate.
Caspase Activity Assays
A highly specific alternative to Western blots or single-substrate caspase assays, these kits contain Capture Antibody, Precursor Capase-3, and DEVD-AFC. The target active caspase (ex. Caspase-8) is bound by Capture Antibody. Captured caspase cleaves and activates Precursor Caspase-3 which in turn cleaves the fluorescent substrate DEVD-AFC to generate signal.
Phosphatase Activity Assays
A sensitive and highly selective alternative to single substrate phosphatase assays, these kits contain Capture Antibody, phospho-peptide substrate, and Malachite green for detecting inorganic phosphate. The target phosphatase is bound by the Capture Antibody. The captured phosphatase liberates phosphate from the phospho-peptide substrate and the free phosphate is measured by a change in color of Malachite green.
Receptor Tyrosine Kinase Assays
A rapid and more sensitive alternative to Western blots, Phospho Receptor Tyrosine Kinase (RTK) kits include Capture Antibody, Detection Antibody-HRP Conjugate, and Tyrosine-Phosphorylated Positive Control. In this assay total RTK protein is captured using immobilized Capture Antibody and tyrosine phosphorylated RTK is detected using an anti-phosphotyrosine Detection Antibody-HRP Conjugate (substrate sold separately, Catalog # DY999 ). Total RTK kits include Capture Antibody, Standard, Biotinylated Detection Antibody, and Streptavidin-HRP (substrate sold separately, Catalog # DY999 ). Total RTK protein is sandwiched between an immobilized Capture Antibody and a Biotinylated Detection Antibody. Signal is generated using Streptavidin-HRP and substrate.
Transcription Factor Activity Assays
A reliable alternative to electrophoretic mobility shift assays (EMSA), these kits contain Capture Antibody, Biotinylated double-stranded Oligonucleotide, Unlabeled double-stranded Oligonucleotide, and Streptavidin-HRP. The target active transcription factor binds Biotinylated double-stranded Oligonucleotide in solution. The complex is bound by immobilized Capture Antibody and detected using Streptavidin-HRP and substrate (substrate sold separately, Catalog # DY999 ). Several DuoSet IC transcription factor assays can be converted to measure total (active and inactive) transcription factor (see product inserts for details).
Immunoprecipitation (IP)-Kinase Assays
IP Kinase assays offer a highly specific and highly sensitive way to measure endogenous kinase activity. These assays are simple to perform and provide rapid, quantitative results. Available in a single 96 well plate pack, each kit provides a Kinase-specific Antibody conjugated to agarose beads, HRP-conjugated Detection Reagent, Streptavidin-coated Microplate, Biotinylated Kinase Substrate, Biotinylated Phospho-peptide Control, Substrate Solution, and Protocol Manual. The captured kinase phosphorylates the provided substrate, and phosphorylated substrate is assessed using colorimetric detection.
* IP-Kinase Assays are only available as 1-96 well plate packs
References
- Elevated Expression Level of Survivin Protein in Soft-tissue Sarcomas is a Strong Independent Predictor of Survival. Kappler, M. et al. (2003) Clin. Cancer Res. 9:1098.
R&D Catalog Numbers: AF886; DYC647