BCL-2 (Phospho-Thr56)Colorimetric Cell-Based ELISA Kit

This colorimetric cell-based ELISA kit from Assay Genie is designed for the precise quantification of BCL-2 phosphorylation at threonine 56 in cell samples. The assay offers high sensitivity and specificity, delivering accurate and reproducible results for research applications studying cell signaling pathways and exploring potential targets for drug development. BCL-2 is a key protein involved in regulating apoptosis, or programmed cell death, and its phosphorylation at threonine 56 has been linked to various diseases including cancer and neurodegenerative disorders.

The kit uses a convenient, lysate-free, high-throughput indirect ELISA format. BCL-2 (Phospho-Thr56) is captured by specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. The HRP enzyme conjugated to the secondary antibody catalyzes a colorimetric reaction measured at 450 nm upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing the target absorbance values, and the Crystal Violet whole-cell staining method can be used to determine cell density, allowing results to be normalized to cell amounts so that plating differences can be adjusted.

Intended for adherent cell samples with human reactivity and a dynamic range of > 5000 cells, this Cell-Based ELISA can detect BCL-2 protein phosphorylation and expression profiles in cultured cells. It can be used to measure the relative amounts of phosphorylated BCL-2 as well as to screen the effects of various treatments, inhibitors such as siRNA or chemicals, or activators on BCL-2 phosphorylation. BCL-2 is an antiapoptotic member of the Bcl-2 family that regulates cell death by controlling mitochondrial membrane permeability, inhibiting caspase activity either by preventing the release of cytochrome c from the mitochondria or by binding to the apoptosis-activating factor (APAF-1), with phosphorylation by JNKs potentially increasing its antiapoptotic functions.