Myc (phospho Thr58) Cell Based ELISA Kit

This Cell-Based ELISA kit from Antibodies.com is an Enzyme-Linked Immunosorbent Assay designed for the indirect in vitro quantitative determination of c-Myc levels in adherent and suspension cells. It allows for the detection of Myc (phospho Thr58) and the effects that certain stimulation conditions have on Myc (phospho Thr58) expression in different cell lines. Qualitative determination of Myc (phospho Thr58) concentration is achieved by an indirect ELISA format, in which the target is captured by Anti-Myc (phospho Thr58) Antibodies and detected by HRP-conjugated secondary antibodies. Through this binding, the HRP enzyme conjugated to the secondary antibody can catalyze a colorimetric reaction upon substrate addition.

The assay is reactive with Human (Thr58), Mouse (Thr58), and Rat (Thr58), with no reported cross-reactivity. Detection is colorimetric, performed on a Tissue Culture Treated Microplate platform, with an assay range of > 5000 Cells and an assay time of 4 hours 30 minutes. Due to the qualitative nature of the kit, multiple normalization methods are described: an Anti-GAPDH Antibody is included to serve as an internal positive control in normalizing target absorbance values; following the colorimetric measurement of HRP activity, the Crystal Violet whole-cell staining method is used to determine cell density, allowing absorbance values to be normalized to cell amounts and adjusting for plating differences; and an Anti-Myc Antibody is provided for normalization, where absorbance values for the non-phosphorylated target can be used to normalize values for the phosphorylated target.

The kit is suitable for use with adherent and suspension cells as sample types. Storage is at + 4°C, and the kit is stable for 3-6 months. The modification detected is phospho Thr58, and the protein target is c-Myc. The assay type is cell-based (qualitative).