Catalogue

Background
There are two known isoforms of MiTF differing by 66 amino acids at the NH2 terminus. Shorter forms are expressed in melanocytes and run as two bands at 52kDa and 56kDa, while the longer Mi form runs as a cluster of bands at 60-70kDa in osteoclasts and in B16 melonoma cells (but not other melanoma cell lines), as well as mast cells and heart. It reacts with both melanocytic as well as the non- melanocytic isoforms of MiTF. This Ab does not cross-react with other b-HLH-ZIP factors by DNA mobility shift assay. Mi is a basic helix-loop-helix-leucin zipper (b-HLH-ZIP) transtripotion factor implicated in pigmentation, mast cells and bone development. The mutation of MiTF causes Waardenburg Syndrome type II in humans. In mice, a profound loss of pigmented cells in the skin eye and inner ear results, as well as osteopetrosis and defects in natural killer and mast cells. These melanocyte isoforms have been shown by two dimensional tryptic mapping to differ in c-Kit-induced phosphorylation. Osteopetrotic rat strain harbors a large genomic deletion encompassing the 3 prime end of TF including most of the b-HLH-ZIP region. Osteoclasts from these animals lack MiTF protein in contrast to wild-type rat, mouse, and human osteoclasts.

Synonyms: MiTF

Source
Hybridoma produced by the fusion of splenocytes from RBF/DnJ mice immunized with an N-terminal fragment of human microphthalmia protein and mouse myeloma NS1 cells.

Product
In Western blotting, it recognizes a doublet of 52-56kDa, identified as serine-phosphorylated and unphosphorylated forms of melanocytic isoforms of microphthalmia (Mi) transcription factor

Product Form: Unconjugated

Formulation: Provided as solution in phosphate buffered saline with 0.08% sodium azide

Purification Method: Protein A/G Chromatography

Concentration: See vial for concentration

Applications
This antibody cocktail of clone C5 and D5 can be used for gel supershift assays, immunoprecipitation (2 µg/mg of protein lysate), Western blotting (1 µg/ml) and immunohistochemistry on frozen and formalin/paraffin tissue sections. Optimal concentration should be evaluated by serial dilutions. In Western blotting, it recognizes a doublet of 52-56kDa, identified as serine-phosphorylated and unphosphorylated forms of melanocytic isoforms of microphthalmia (Mi) transcription factor

Functional Analysis: Western Blotting

Positive Control: 501 Mel human melanoma cells, wild-type human, rat, mouse osteoclast cells

Storage
Product should be stored at -20°C. Aliquot to avoid freeze/thaw cycles

Product Stability: See expiration date on vial

Shipping Conditions: Ship at ambient temperature, freeze upon arrival

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but our company accepts no liability for any inaccuracies or omissions in this information.

References
1. Weilbaecher KN, et. al. Age-resolving osteopetrosis: a rat model implicating microphthalmia and the related transcription factor TFE3. J. Exp.Med. 1998, 187: 775-785 2. Hemesath P, et. al. MAP kinase links the transcription factor microphthalmia to c-Kit signalling in melanocytes. Nature. 1998, 391:298-301

Protein Reference(s)

Database Name: UniProt

Accession Number: O75030

Safety Datasheet(s) for this product: