MITF Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

MITF (microphthalmia-associated transcription factor) is a melanocytic nuclear protein that contains basic helix-loop-helix (HLH) and leucine zipper (LZ) domains. These protein motifs are frequently observed in other transcription factors and are particularly common to members of the Myc family. MITF can directly associate with DNA as a homodimer. It is required for the development and differentiation of melanocytes. Its expression is upregulated by cAMP and cAMP dependent pathways. MITF activates several different gene promoters by binding to their E-boxes. Tyrosinase, TRP-1 and TRP-2 are pigment synthesis genes activated by MITF. When MITF is phosphorylated on Serine 73 (via the MAPK pathway), it associates with coactivators of the p300/CBP family and enhances transcription. MITF has several isoforms including MITF-M which is specifically expressed in melanocytes. In Mitfdeficient mice there is a complete absence of melanocytes.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-09070

• MITF Antibodies
• MITF ELISA Kits
• MITF Lysate
• MITF Protein