Claudin 2 Colorimetric Cell-Based ELISA Kit

Assay Genie provides a cell-based ELISA kit for the quantitative analysis of Claudin-2 protein levels in cell samples. Designed as a high-throughput, lysate-free and sensitive assay, the kit detects the Claudin 2 protein expression profile in cultured cells and can be used to measure relative amounts of Claudin 2 as well as to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals) or activators on Claudin 2. The assay uses an indirect ELISA format: Claudin 2 is captured by Claudin 2-specific primary antibodies while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, and the conjugated HRP enzyme catalyses a colorimetric reaction upon substrate addition.

Because the Cell-Based ELISA provides qualitative determination, two normalization strategies are described. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalizing target absorbance values, and following the colorimetric measurement of HRP activity the Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalized to cell amounts so that plating differences can be adjusted. The kit is reactive with human and mouse samples and is intended for adherent cells. Detection is colorimetric, read at 450 nm, with a dynamic range of > 5000 cells, and storage stability is given as 4 °C for 6 months.

Claudin-2 is a tight-junction protein that plays a major role in tight junction-specific obliteration of the intercellular space through calcium-independent cell-adhesion activity (UniProt ID: P57739; molecular weight 25 kDa). Dysregulation of Claudin-2 has been implicated in inflammatory bowel disease, cancer and renal disorders, making it a valuable target for basic and therapeutic research into epithelial permeability and related disease processes.