Mouse Microphthalmia Transcription Factor (MiTF) Monoclonal Antibody | anti-MiTF antibody

Microphthalmia Transcription Factor (MiTF)

Cat. Num. AAA395097

• Gene Names: MITF; MI; WS2; CMM8; WS2A; bHLHe32
• Reactivity: Human, Mouse, Rat
• Applications: Gel Super Shift Assay, Western Blot, Immunoprecipitation, Immunohistochemistry
• Purity: Protein A/G Chromatography
• Synonyms: Microphthalmia Transcription Factor (MiTF); Monoclonal Antibody; MiTF; anti-MiTF antibody

• Host: Mouse
• Reactivity: Human, Mouse, Rat
• Clonality: Monoclonal
• Isotype: IgG1
• Clone Number: C5
• Purity/Purification: Protein A/G Chromatography
• Form/Format: Provided as solution in phosphate buffered saline with 0.08% sodium azide
• Sequence Length: 419

• Applicable Applications for anti-MiTF antibody: Gel Supershift (GS), Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC) Frozen/Paraffin
• Immunogen: 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.
• Positive Control: 501 Mel human melanoma cells, wild-type human, rat, mouse osteoclast cells
• Preparation and Storage: Product should be stored at -20 degree C. Aliquot to avoid freeze/thaw cycles

Immunohistochemistry (IHC)

(Immunohistochemical staining using microphthalmia antibody on formalin fixed, paraffin embedded human)

Related Product Information for anti-MiTF antibody

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. 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? half of MiTF 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.

Product Categories/Family for anti-MiTF antibody

Monoclonal Antibody; Transcription Factors

NCBI and Uniprot Product Information

• NCBI GI #: 4557755
• NCBI GeneID: 4286
• NCBI Accession #: NP_000239.1
• NCBI GenBank Nucleotide #: NM_000248.3
• UniProt Accession #: O75030; Q14841; Q9P2V0; Q9P2V1; Q9P2V2; Q9P2Y8; B4DJL2; D3K197; E9PFN0
• Molecular Weight: 52-56kDa doublet
• NCBI Official Full Name: microphthalmia-associated transcription factor isoform 4
• NCBI Official Synonym Full Names: microphthalmia-associated transcription factor
• NCBI Official Symbol: MITF
• NCBI Official Synonym Symbols: MI; WS2; CMM8; WS2A; bHLHe32
• NCBI Protein Information: microphthalmia-associated transcription factor; class E basic helix-loop-helix protein 32
• UniProt Protein Name: Microphthalmia-associated transcription factor
• Protein Family: Microphthalmia-associated transcription factor
• UniProt Gene Name: MITF
• UniProt Synonym Gene Names: BHLHE32; bHLHe32
• UniProt Entry Name: MITF_HUMAN

NCBI Description

This gene encodes a transcription factor that contains both basic helix-loop-helix and leucine zipper structural features. It regulates the differentiation and development of melanocytes retinal pigment epithelium and is also responsible for pigment cell-specific transcription of the melanogenesis enzyme genes. Heterozygous mutations in the this gene cause auditory-pigmentary syndromes, such as Waardenburg syndrome type 2 and Tietz syndrome. Alternatively spliced transcript variants encoding different isoforms have been identified. [provided by RefSeq, Jul 2008]

Uniprot Description

Function: Transcription factor that regulates the expression of genes with essential roles in cell differentiation, proliferation and survival. Binds to symmetrical DNA sequences (E-boxes) (5'-CACGTG-3') found in the promoters of target genes, such as BCL2 and tyrosinase (TYR). Plays an important role in melanocyte development by regulating the expression of tyrosinase (TYR) and tyrosinase-related protein 1 (TYRP1). Plays a critical role in the differentiation of various cell types, such as neural crest-derived melanocytes, mast cells, osteoclasts and optic cup-derived retinal pigment epithelium. Ref.11 Ref.19

Subunit structure: Efficient DNA binding requires dimerization with another bHLH protein. Binds DNA in the form of homodimer or heterodimer with either TFE3, TFEB or TFEC. Interacts with KARS. Identified in a complex with HINT1 and CTNNB1. Ref.12 Ref.19

Subcellular location: Nucleus.

Tissue specificity: Isoform M is exclusively expressed in melanocytes and melanoma cells. Isoform A and isoform H are widely expressed in many cell types including melanocytes and retinal pigment epithelium (RPE). Isoform C is expressed in many cell types including RPE but not in melanocyte-lineage cells. Isoform Mdel is widely expressed in melanocytes, melanoma cell lines and tissues, but almost undetectable in non-melanoma cell lines. Ref.3

Post-translational modification: Phosphorylation at Ser-405 significantly enhances the ability to bind the tyrosinase promoter. Phosphorylated at Ser-180 and Ser-516 following KIT signaling, trigerring a short live activation: Phosphorylation at Ser-180 and Ser-516 by MAPK and RPS6KA1, respectively, activate the transcription factor activity but also promote ubiquitination and subsequent degradation by the proteasome. Ref.10 Ref.11Ubiquitinated following phosphorylation at Ser-180, leading to subsequent degradation by the proteasome. Deubiquitinated by USP13, preventing its degradation. Ref.10

Involvement in disease: Waardenburg syndrome 2A (WS2A) [MIM:193510]: WS2 is a genetically heterogeneous, autosomal dominant disorder characterized by sensorineural deafness, pigmentary disturbances, and absence of dystopia canthorum. The frequency of deafness is higher in WS2 than in WS1.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.20Waardenburg syndrome 2, with ocular albinism, autosomal recessive (WS2-OA) [MIM:103470]: A disorder characterized by the association of features typical of Waardenburg syndrome type 2 with ocular albinism. Patients manifest reduced visual acuity, albinotic fundus, deafness, hypomelanosis.Note: The disease is caused by mutations affecting the gene represented in this entry.Tietz syndrome (TIETZS) [MIM:103500]: Autosomal dominant disorder characterized by generalized hypopigmentation and profound, congenital, bilateral deafness. Penetrance is complete.Note: The disease is caused by mutations affecting the gene represented in this entry. Ref.21Melanoma, cutaneous malignant 8 (CMM8) [MIM:614456]: A malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.Note: Disease susceptibility is associated with variations affecting the gene represented in this entry.

Sequence similarities: Belongs to the MiT/TFE family.Contains 1 bHLH (basic helix-loop-helix) domain.