TNFSF11 Colorimetric Cell-Based ELISA

This Colorimetric Cell-Based ELISA Kit from Assay Genie is designed for the accurate measurement of TNFSF11 levels in cell lysates and tissue homogenates. TNFSF11, also known as RANKL (Receptor Activator of Nuclear Factor Kappa-B Ligand), is a critical regulator of bone metabolism, playing a pivotal role in cell differentiation, survival, and bone resorption. The kit has been validated for use with Human and Mouse samples and is configured for adherent cell workflows.

The assay uses an indirect ELISA format, in which TNFSF11 is captured by target-specific primary antibodies and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. The conjugated HRP enzyme then catalyses a colorimetric reaction upon substrate addition, read at 450 nm. The kit supports high throughput screening and can be used to measure the relative amounts of TNFSF11 in cultured cells as well as to screen for the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on TNFSF11. Because the assay is qualitative, two normalization methods are supplied: a monoclonal antibody specific for human GAPDH serves as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining method can be used to determine cell density after the colorimetric measurement.

The kit is recommended for storage at 4°C, with a stated stability of six months under those conditions, and supports a dynamic range of greater than 5000 cells. Research applications include the study of bone-related disorders such as osteoporosis and osteopetrosis, as well as inflammatory conditions such as rheumatoid arthritis, where imbalances in TNFSF11 levels have been linked to disease.