Product Name:
MITF-pS414
Product Number:
ab-pn626
Target Full Name: Microphthalmia-associated transcription factor
Target Alias: BHLHe32; BW; Homolog of mouse microphthalmia; MI; VIT; WS2A
Product Type Specific: MITF phosphosite-specific antibody
Antibody Code: PN626
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S414
Protein UniProt: O75030
Protein SigNET: MITF
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human MITF
Antibody Immunogen Sequence: TGLA(pS)PDL(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues T410 to D417. The effect of S414 phosphorylation is unclear. This is the major in vivo phosphorylation sites in MITF (≥24 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). MITF is known to be phosphorylated at this site in vitro by p38a MAPK (MAPK14).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rhesus Macaque | Mouse | Chicken.
Scientific Background: MITF (Microphthalmia-associated transcription factor) is a transcription factor in the MiT/TFE family. It is a master regulator protein essential for the development, survival, and function of melanocytes, osteoclasts, mast cells and optic cup-derived retinal pigment epithelium. As a basic helix-loop-helix leucine zipper (bHLH-Zip) transcription factor, it acts as a dimer (homo- or heterodimer) and undergoes post-translational modifications, including phosphorylation (by MAPK, MTOR, GSK3) and SUMOylation. It binds to M-boxes (5'-TCATGTG-3') and symmetrical DNA sequences (E-boxes) (5'-CACGTG-3') found in the promoter of pigmentation genes, such as Bcl-2, tyrosinase (TYR) and tyrosinase-related protein 1 (TYRP1). It controls pigment production (melanin) and regulates genes involved in cell cycle and apoptosis. It is involved in the cellular response to amino acid availability by acting downstream of MTOR. In the presence of nutrients, MITF phosphorylation by MTOR promotes its inactivation. Upon starvation or lysosomal stress, inhibition of MTOR induces MITF dephosphorylation, resulting in transcription factor activity. 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 poorly in non-melanoma cell lines. Loss-of-function mutations in MITF cause Waardenburg syndrome type 2A and Tietz syndrome (deafness and pigment loss), while amplification or specific mutations act as an oncogene in melanoma. This description may include information annotated by UniProt and/or Google AI.