详细说明
Species Reactivity
Human
Specificity
Detects human MITF in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human MITF
Val119-Lys289
Accession # O75030Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
1 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Immunohistochemistry | 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 # ). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # ) and counterstained with hematoxylin (blue). View our protocol for . |
Western Blot | 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 # ). A specific band was detected for MITF at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Preparation and Storage
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping
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.
Background: MITF
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.
Long Name:
Microphthalmia-associated Transcription Factor
Entrez Gene IDs:
4286 (Human); 17342 (Mouse); 25094 (Rat)
Alternate Names:
bHLHe32; bHLHe32MI; Class E basic helix-loop-helix protein 32; MI; microphthalmia-associated transcription factor; MITF; Waardenburg syndrome, type 2A; WS2A