Human Plakophilin 1 ELISA Kit

A sandwich double-antibody ELISA from Assay Genie designed for quantitative measurement of human plakophilin-1 (also referred to as PKP1, B6P, or Band 6 protein) using colorimetric detection. The assay is reactive with human samples and accepts serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Sensitivity is 0.188 ng/mL with a working range of 0.313–20 ng/mL, and kit components are intended for storage at 2–8°C for 12 months.

Validation data are reported for serum, EDTA plasma, and heparin plasma (n=5 each). Linearity at 1:2, 1:4, and 1:8 dilutions ranged 86–105%, 84–98%, and 82–97% in serum; 92–97%, 87–101%, and 82–88% in EDTA plasma; and 86–97%, 84–95%, and 81–100% in heparin plasma. Spike-and-recovery averages were 94% in serum (range 87–103%), 95% in EDTA plasma (range 88–102%), and 94% in heparin plasma (range 87–101%).

The protocol uses a pre-coated plate: standards, samples, and controls are loaded and incubated at 37°C for 90 minutes, followed by a biotinylated detection antibody (37°C, 60 minutes), HRP-streptavidin binding (37°C, 30 minutes), TMB substrate incubation in the dark for 10–20 minutes, stop solution, and immediate absorbance reading at 450 nm. Sample-preparation guidance is provided for serum (clot, 1000 × g, 20 minutes), plasma (anticoagulant tube, 1000 × g, 15 minutes at 2–8°C), tissue homogenate in PBS with protease inhibitors, cell culture supernatant (2500 rpm, 5 minutes), and cell lysates. Plakophilin-1 is involved in cell–cell adhesion and epithelial tissue integrity, with dysregulation linked to skin disorders, cardiomyopathies, and certain cancers, supporting its relevance as a biomarker in cell-biology research.