SpectraMax Paradigm Multi-Mode Microplate Reader

Kinase/Phosphatase Assays

Kinases are one of the most critical targets in drug discovery today. These enzymes are key components in cellular signaling pathways, and disruption to their function causes a variety of diseases, such as metabolic diseases, certain cancers, and cardiac disease. The functionally related phosphatases and phosphodiesterases are also important screening targets.

Read Application Note 

Western Blot Detection

Western blotting is among the most common methods employed for the detection and quantitation of specific proteins. In this method, protein sample, e.g. from a cell lysate, is first separated by size on an SDS-PAGE gel. Proteins are then transferred to a nitrocellulose or PVDF membrane, which is probed with antibody specific to the protein of interest. Various techniques are used to detect proteins on western blot membranes including fluorescence and chemiluminescence.

Read Application Note 

HTRF

HTRF®, an assay technology developed by Cisbio, combines standard FRET (fluorescence resonance energy transfer) with time-resolved measurement of fluorescence, eliminating short-lived background fluorescence. HTRF’s applications include assessment of small phosphorylated peptides, immunoassays for quantifying large glycoproteins, and indirect detection of tagged complexes such as CD28/CD86 via secondary antibodies. These homogeneous assays do not require wash steps and are amenable to small-volume microplate formats used in high-throughput screening.
(This assay is suitable for SpectraMax® iD5 and SpectraMax® Paradigm readers.)

Read Application Note 

AlphaScreen

AlphaScreen® Technology is a homogeneous, high-throughput method for detecting intramolecular binding. Donor and acceptor beads are attached to biomolecules, and when the beads are brought into proximity (via binding), a cascade of chemical reactions occurs, amplifying a signal that is detected on the SpectraMax® i3x and Paradigm® Multi-Mode Microplate Readers.