Collagen I alpha2 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit enables the qualitative determination of Collagen I alpha2 (COL1A2) protein expression in cultured cells. Using an indirect ELISA format, the target is captured by Collagen I alpha2-specific primary antibodies while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition. The assay is lysate-free, high throughput and sensitive, and can be used to measure relative Collagen I alpha2 levels as well as to screen the effects of various treatments, inhibitors (such as siRNA or chemicals) or activators.

Reactivity is human and the kit is intended for adherent cells, with a suggested seeding of 200 µL of 20,000 adherent cells per well of a 96-well plate. Detection is colorimetric at 450 nm, with a dynamic range of > 5000 Cells. Storage stability is 4 °C for 6 Months. An anti-GAPDH monoclonal antibody is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step may be used after HRP measurement to determine cell density and adjust absorbance values for plating differences.

The target is encoded by the COL1A2 gene (Gene ID 1278; UniProt ID P08123; OMIM 120160) and is a fibril-forming component of the extracellular matrix with roles in tissue development, repair and remodelling. Abnormal levels have been linked to fibrosis, arthritis and cancer. Defects in COL1A2 are associated with Ehlers-Danlos syndrome type 7B, osteogenesis imperfecta types 1–4, and the autosomal recessive cardiac valvular form of Ehlers-Danlos syndrome.