Human and Mouse Phospho-FOXO3 (S253) PharmaGenie ELISA Kit (SBRS1831)

A colourimetric phosphorylation ELISA from Assay Genie is designed to detect phospho-FOXO3 at serine 253. The kit targets the FOXO3 gene product, also known as FKHRL1 and FOXO3A (Forkhead box protein O3; AF6q21 protein; Forkhead in rhabdomyosarcoma-like 1), with Uniprot references O43524 and Q9WVH4 and Gene ID 2309. It is validated for use with human samples, including serum, plasma and cell lysates, as demonstrated in experiments with HeLa cells treated with Calyculin A and analysed by both the phosphoELISA and Western blot.

Supplied components include a pre-coated strip microplate, wash buffer, anti-phospho antibody, HRP-conjugated secondary antibody, assay diluent, TMB one-step substrate, stop solution, lysis buffer and a positive control sample. The assay protocol involves adding 100 ul of sample or positive control per well, incubating 2.5 h at room temperature or overnight at 4 °C, followed by addition of 100 ul of prepared primary antibody with a 1 h room-temperature incubation, then 100 ul of prepared 1X HRP-streptavidin with another 1 h incubation, 100 ul of TMB one-step substrate reagent with a 30 min incubation, 50 ul of stop solution, and immediate absorbance reading at 450 nm. The kit should be stored at -20 °C and used within 6 months of the date of shipment.