ST5 Colorimetric Cell-Based ELISA

This Cell-Based ELISA Kit from Assay Genie is designed for the detection and quantification of the ST5 protein in cell samples. Reactive with Human and Mouse samples, it is suited for use with adherent cells and supports a variety of applications including the study of protein expression, signaling pathways, and drug efficacy in cell-based research.

The assay uses a colorimetric detection method read at 450 nm, with a stated dynamic range of > 5000 Cells. It is supplied in a 96-Well Microplate format and should be stored at 4°C for up to 6 months. The procedure is lysate-free and high throughput, following an indirect ELISA format in which ST5 is captured by ST5-specific primary antibodies and detected using HRP-conjugated secondary antibodies. A monoclonal anti-GAPDH antibody is included to serve as an internal positive control, and Crystal Violet whole-cell staining may be used to determine cell density for normalization of absorbance values.

ST5, also known as Suppression of tumorigenicity 5 protein or DENND2B, is a guanine nucleotide exchange factor (GEF) that may activate RAB9A and RAB9B. The protein has a molecular weight of 70,245 Da and is encoded by a gene located at chromosomal position 11p15 (Gene ID: 6764; UniProt ID: P78524). The workflow involves seeding adherent cells, fixation, quenching, blocking, overnight primary antibody incubation at 4°C with ST5-specific antibodies, secondary antibody incubation, addition of Ready-to-Use Substrate, and immediate reading of OD at 450 nm following Stop Solution addition.