The In-Cell Western™ Assay is an immunocytochemical assay that uses near-infrared fluorescence to detect and quantify proteins in fixed cells. Detecting proteins in their cellular context increases quantification precision. Proteins in fixed, cultured cells are detected directly in microplates, which yields higher throughput compared to Western blotting and eliminates typical Western blotting steps such as cell lysate preparation, electrophoresis, and membrane transfer. Using the In-Cell Western Assay kits, the cost per well for secondary screening is reduced to a fraction of the cost of typical screening methods. Watch an introductory webinar to In-Cell Western Assays.
The In-Cell Western™ Assay Kits provide antibodies, blocking buffer, and stains for forty 96-well plates or ten 384-well plates. Sapphire700™ and DRAQ5™ protein stains are included to normalize for well-to-well differences in cell number. Using protein stains reduces the cost per assay compared to performing the assay using two secondary antibodies. Any potential interference caused by using two antibodies is also eliminated.
Additional Resources:
In-Cell Western Assay Protocol: Complete Apoptosis Assay Example Detailing the Seeding, Induction, and Detection of the HeLa Cellular Response to Anisomycin Treatment
Odyssey Publications List: In-Cell Western Assays, Vol 2, Winter 2012
Other In-Cell Western Assay Protocols are available on our Technical Resources Library. Visit our In-Cell Western Application Pages for more examples of how researchers have used these immunocytochemical assays.
