The Bio-Plex® phospho 3-plex immunoassay is a 1 x 96-well multiplex set that contains a vial of coupled magnetic beads, a vial of detection antibodies, and cell lysates for the detection of phosphorylated proteins in lysates derived from cell culture or tissue samples. This assay detects:
- p70 S6 kinase
- STAT3 (Tyr705)
The assays in this multiplex set are based on 5.6 µm nonmagnetic beads. For the multiplexing capabilities of specific assay combinations, see bulletin 5483.
Bio-Plex offers total target singleplex and phosphoprotein singleplex set assays. Bio-Plex total target assays measure the total abundance of a target protein, regardless of its phosphorylation state, and can be used to report the total concentration of a protein. Bio-Plex phosphoprotein assays measure the concentration of the phosphorylated protein, only. In cases where phosphoprotein and total protein assays are offered, they must be run in different wells due to shared bead regions. Using the multiplex format, the profiles of multiple phosphorylated proteins involved in cell signaling pathways can be detected using only 25 µl of lysate.
- Highly specific antibodies exclusively developed and validated by Cell Signaling Technology, Inc.
- 96-well filter-plate format which yields significantly higher throughput than traditional methods
- Higher specificity sandwich format which minimizes cross reactivity with other cellular proteins
- Direct measurement of kinases that have been activated within the cell
- Low sample volume requirement
- Minimal hands-on times
Additional Necessary Reagents
In addition to this phosphoprotein assay, the following items are needed to perform a Bio-Plex signal transduction assay:
- Bio-Plex phosphoprotein detection reagent kit (#171-304004, 1 x 96-well) — includes assay reagents and instructions required to prepare samples and acquire data
- Bio-Plex cell lysis kit (#171-304011, 1 x 96-well) — includes buffers and factors essential for the optimal lysis of cell culture or tissue samples
- Bio-Plex 200 system
- Bio-Plex 3D system
- Bio-Plex Pro™ II wash station