E2F1 Colorimetric Cell-Based ELISA Kit

Designed for the accurate detection of E2F1 levels in cell lysates, tissue homogenates, and other biological samples, this colorimetric cell-based ELISA kit from Assay Genie offers a convenient, lysate-free, high-throughput and sensitive assay for measuring the relative expression profile of E2F1 protein in cultured adherent cells, and can be used to screen for the effects that various treatments, inhibitors such as siRNA or chemicals, or activators have on E2F1.

E2F1 is a key transcription factor that regulates genes involved in cell cycle progression, apoptosis, and differentiation, and its dysregulation has been implicated in various diseases, including cancer. The assay uses an indirect ELISA format in which E2F1-specific primary antibodies capture the target while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, allowing the HRP enzyme to catalyse a colorimetric reaction upon substrate addition that is read at 450 nm for a qualitative determination of E2F1 concentration.

Because the Cell-Based ELISA is qualitative, multiple normalisation methods are supported: a monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and following the colorimetric measurement the Crystal Violet whole-cell staining method may be used to determine cell density so that absorbance values can be normalised to cell amounts to adjust for plating differences. The kit is reactive with human and mouse samples, has a stated dynamic range of > 5000 Cells, and is supplied for storage at 4 °C with a stability of 6 Months.