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Purified anti-MiTF Antibody

Pricing & Availability
Clone
W23160C (See other available formats)
Regulatory Status
RUO
Other Names
Microphthalmia-associated transcriptionfactor, Class E basic helix-loop-helix protein 32, bHLHe32
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
A. W23160C_PURE_MiTF_WB_091725
Whole cell extracts (15 µg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-MiTF (clone W23160C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • A. W23160C_PURE_MiTF_WB_091725
    Whole cell extracts (15 µg total protein per lane) from indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-MiTF (clone W23160C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection reagent. Lane M: Molecular weight marker
  • B. W23160C_PURE_MiTF_IHC-P_091725
    IHC staining with Purified anti-MiTF (clone W23160C) on formalin-fixed paraffin-embedded human melanoma tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated with (panel A) and without (panel B) Purified anti-MiTF (clone W23160C) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (panels C and D). Images were captured with a 40X objective and merged (panels E and F). Scale bar: 50 µm
  • C, W23160C_PURE_MiTF_ICC_091725
    SK-MEL-28 cells (high-expressing cell line, panel A) and HeLa cells (low-expressing cell line, panel D) were fixed with Fixation Buffer (Cat. No. 420801) for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with Purified anti-MiTF (clone W23160C), followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Cells were co-stained with Alexa Fluor® 488 anti-Tubulin-α (Cat. No. 627906) and nuclei were counterstained with DAPI (Cat. No. 422801) (panels B and E). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 63X objective and merged (panels C and F). Scale bar: 50 µm
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648876 25 µg $195
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648877 100 µg $490
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Description

Microphthalmia-associated transcription factor (MiTF) is a basic helix–loop–helix leucine zipper (bHLH-LZ) transcription factor which serves as a master regulator of melanocyte development, function, and survival. MiTF binds DNA as a homodimer or heterodimer and recognizes specific E-box sequences (CANNTG) to regulate transcription. MiTF has multiple isoforms generated via alternative promoter usage and splicing. MiTF contains an N-terminal activation domain, the central bHLH-LZ domain responsible for DNA binding/dimerization, and a C-terminal transcriptional activation/repression domain. MiTF expression and activity are controlled by multiple signaling pathways including MAPK/ERK, WNT/β-catenin, cAMP/PKA, and PI3K/AKT, through transcriptional, post-transcriptional, and post-translational mechanisms such as phosphorylation, sumoylation, and ubiquitination. During development, MiTF is essential for the specification of the neural crest-derived melanocyte lineage. It governs melanoblast migration, proliferation, melanin synthesis, and differentiation into pigmented melanocytes by activating key melanogenic genes such as TYR, TYRP1, DCT and PMEL. In melanoma, MiTF acts as a lineage survival oncogene. Gene amplification or dysregulated signaling leading to increased MiTF activity is observed in subsets of melanoma, enhancing tumor survival and resistance to apoptosis. In humans, MiTF mutations result in various developmental disorders, such as Waardenburg syndrome type II (WS2) and Tietz syndrome.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of C-terminus of human MiTF
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P, ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunohistochemistry, a concentration range of 1.0 - 10.0 µg/mL is suggested. For immunocytochemistry, a concentration range of 2.5 - 10.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Additional Product Notes

For use in immunohistochemistry on formalin-fixed paraffin-embedded tissue (IHC-P), it is recommended to perform antigen retrieval using either Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 Antigen Retrieval Buffer (10X) (Cat. No. 422704).

For use in immunocytochemistry (ICC), it is recommended to fix/permeabilize with either of the following:
-Fixation buffer (Cat. No. 420801) with 0.5 % Triton X-100
-Fixation buffer (Cat. No. 420801) with 100% ice-cold methanol
-100% ice-cold methanol only

Antigen Details

Structure
Canonical MiTF is a 526 amino acid protein with a predicted molecular weight of 58.8 kD. The molecular weight of the other 11 isoforms vary from 40 to 58.1 kD.
Distribution

Located in nucleus

Function
Transcription factor that regulates melanocyte survival, differentiation, and melanosome biogenesis
Interaction
Forms homodimer or heterodimer with TFE3, TFEB, or TFEC
Cell Type
Epithelial cells, Mast cells, Osteoclasts
Biology Area
Neuroscience, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References
  1. Garraway LA, et al. 2005. Cell. 120(3):335-348.
  2. Strub T, et al. 2011. Cancer Cell. 19(6):715-730. 
  3. Laurette P, et al. 2015. eLife. 4:e06857. 
  4. Verfaillie A, et al. 2015. Nat. Cell Biol. 17(2):259-271.

 

Gene ID
4286 View all products for this Gene ID
UniProt
View information about MiTF on UniProt.org

Related FAQs

There are no FAQs for this product.

Other Formats

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Description Clone Applications
Purified anti-MiTF W23160C WB,ICC,IHC-P
Go To Top Version: 1    Revision Date: 09/17/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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