MITF (Phospho-Ser180/73)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the detection of MITF (Microphthalmia-associated transcription factor) phosphorylation at serines 180 and 73 in cell lysates. Configured for adherent cell samples with reactivity against human and mouse targets, the assay provides a qualitative readout by indirect ELISA: target-specific primary antibodies are bound by HRP-conjugated secondary antibodies, which catalyze a colorimetric reaction read at 450 nm. The kit supports a lysate-free, high-throughput workflow for measuring relative levels of phosphorylated MITF in cultured cells, and for screening the effects of treatments, inhibitors (such as siRNA or chemicals) and activators on MITF phosphorylation.

Because the assay is qualitative, two normalization strategies are provided. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalizing target absorbance values, and a Crystal Violet whole-cell staining step allows determination of cell density so absorbance values can be adjusted for plating differences. The stated dynamic range is > 5000 cells.

MITF is a transcription factor containing both basic helix-loop-helix and leucine zipper domains and plays a critical role in the differentiation of neural crest-derived melanocytes, mast cells, osteoclasts, and retinal pigment epithelium. The M-isoform is restricted to melanocytes, while the A-isoform has a broader expression pattern. Phosphorylation at serines 180 and 73 regulates MITF activity, making this phospho-specific assay relevant to research on melanoma progression and pigmentation-related disorders.