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PathHunter® Secretion Detection Kit
The PathHunter® Secretion Detection Kit is for detecting and measuring ProLabel (PL) or ProLink (PK)-tagged recombinant proteins in whole cells following secretion into cell culture media. Using this kit formulation, cell membrane integrity is maintained after the addition of detection reagents. The reagents can be added directly into the assay plate without the need to aspirate and remove the conditioned media to another plate for detection.
Product Features
Simple, homogeneous assay for detecting secretion of either ProLabel or
ProLink-tagged recombinant protein expression in cells
Non-lysing assay buffer allows addition of detection reagents directly to the
assay plate – No need to move conditioned media to another plate
How does the PathHunter® Secretion Detection Kit works
The simple procedure involves addition of EA Reagent and the CL detection reagents directly into the wells of the assay plate. The PL/PK tag fused to the recombinant protein is a small fragment of β-galactosidase that complements with the larger EA fragment within the well, reconstituting active enzyme. The active β-galactosidase hydrolyzes substrate on the membrane, releasing a chemiluminescent signal proportional to the amount of tagged protein present.
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Figure 1. EFC activity of PathHunter® Cell Lines
PathHunter® HEK 293 IL-10-PL Cells were plated in a 96-well plate at 20,000 cells/well and treated
with the known inhibitor, Brefeldin A (Sigma; B7651) for 24 hours. Signal was detected using the
PathHunter® Secretion Detection Kit (93-0237) and the recommended protocol.
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