MITF in Human Skin. MITF was detected in paraffin-embedded sections of human skin using Goat Anti-Human MITF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5769) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human MITF by Western Blot. Western blot shows lysates of Bowes human melanoma cell line and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human MITF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5769) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for MITF at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
MITF (Microphthalmia-associated transcription factor) is a member of the MiT/TFE family of molecules. Although it has a predicted MW of 58 kDa, it runs anomalously in SDS-PAGE at 57‑66 kDa. MITF-A1 is found in melanocytes and RPEs where it regulates melanin synthesis via tyrosinase and TRP-1 gene expression. Human MITF-A1 is 526 amino acids (aa) in length. It contains a bHLH DNA binding region (aa 309‑369) and a Leu-zipper domain (aa 374‑395). There are three phosphorylation sites at Ser54/405/414, and two SUMOylation sites at Arg289/423. MITF-A1 acts as either a homodimer, or heterodimer with TFE3, TFEB or TFEC. Multiple splice variants exist, with each variant expressing one of two isoforms that are defined by the presence (#1), or absence (#2), of aa 294‑299. One variant has a deletion of aa 139‑194, a second has an 11 aa substitution for aa 1‑118, and three others show variable substitutions over aa 1‑35. Over aa 119‑289, human MITF-A1 shares 96% aa identity with mouse MITF-A1.
Microphthalmia-associated Transcription Factor
Entrez Gene IDs:
4286 (Human); 17342 (Mouse); 25094 (Rat)
bHLHe32; bHLHe32MI; Class E basic helix-loop-helix protein 32; MI; microphthalmia-associated transcription factor; Waardenburg syndrome, type 2A; WS2A
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