Revvity, 62GSTPEG, HTRF GST Check Kit, 500 Assay Points

The kit enables the rapid detection of GST tagged fusion proteins at each step of protein production and purification. It can either be used to ensure that the GST motif of a fusion protein is accessible to the anti-GST antibody, or to approximate the concentration of a GST-tagged protein. Calibrated using the appropriate protein, the assay is very accurate and compatible with the SOPs. It can also be used to eliminate false positives. HTRF assays offer many advantages over other technologies: Homogeneous add-and-read format No wash steps Low background Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates Stable signal, providing flexibility in time of readout or size of assays
Assay principle As shown in the graph, GST peptide labeled with XL665 is detected by anti GST Cryptate conjugate (anti-tag Cryptate). The GST-tagged fusion protein (Tag fusion protein) to be ascertained competes with the GST-peptide-XL665 conjugate (Tag-XL665) for binding to the anti-GST-Cryptate conjugate. The specific signal is inversely proportional to the concentration of GST-tagged fusion protein. Assay protocol The example on the right describes the protocol using a 20 µL final assay volume. Dispense 10 µL of standard or of GST-tagged protein of interest into a well, with 5 µL of GST Cryptate conjugate and 5 µL of GST-XL665 conjugate. Incubate for 2 hours at RT and read.