Human Phosphotyrosine Erb-B2 PharmaGenie ELISA Kit (SBRS1815)

Assay Genie's PharmaGenie phosphorylation panels include a lyophilized positive control derived from A431 cells stimulated with EGF at 37°C for 20 minutes, providing a built-in reference for verifying capture and detection reagent activity on every run. Reconstituted positive control lysate should be stored at -70°C to preserve the phospho-ErbB2 signal, while unused microplate strips return to the foil pouch with desiccant and reseal at -20°C. The kit also supplies a dedicated lysis buffer intended to be supplemented with fresh protease and phosphatase inhibitors immediately before cell harvest; cleared supernatant is recovered by microcentrifugation at 13,000 rpm for 10 minutes on ice, aliquoted to avoid repeated freeze-thaw cycles, and either used the same day or snap-frozen at -80°C. The full workflow finishes in roughly four hours at room temperature: a 2.5 hour sample binding step, a 1 hour biotinylated anti-phosphotyrosine incubation, a 45 minute HRP-streptavidin step, and a 30 minute TMB substrate development carried out in the dark before a 50 µL stop solution is added. The capture antibody pair is specific to the human ErbB2 sequence (UniProt P04626, Gene ID 2064) and does not detect the unphosphorylated receptor, so signal changes directly reflect kinase-driven tyrosine phosphorylation rather than total ErbB2 expression levels.