Anti-MITF antibody, rabbit polyclonal, ChIP grade
Product code73-108Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from rabbit antiserum.

Immunogen

Recombinant full-size human Mitf (isoform M) protein with His tag

IsotypeRabbit IgG

Reactivity

Human, mouse, chicken and Xenopus Mitf. This antibody recognizes MITF-M,
MITF-A, MITF-C, MITF-H and other MITF isoforms.

Special notes

Validation: Specificity of the antibody reactivity to Mitf was validated withsiRNAApplication

1. Western blotting (1/1,000-1/3,000)
2. Immunohistochemistry ( 1/300 ~ 1/1,000).
3. Immunofluorescence staining
4.ChIP (1/200)

Background

MITF (Microphthalmia-associated transcription factor) is a transcription factor
that contains both basic helix-loop-helix and leucine zipper structural features.
It plays a critical role in the differentiation of various cell types such as neuralcrest-derived melanocytes, mast cells, osteoclasts and optic cup-derived retinalpigment epithelium.
Mutations in Mitf cause auditory-pigmentary syndromes, such as Waardenburg
syndrome type 2 and Tietz syndrome. Alternatively spliced transcript variants
encoding different isoforms have been identified.

Data Link

UniProtKB Human: O75030 (MITF_HUMAN),
Mouse: Q08874 (MITF_MOUSE), Chicken: O73871 (O73871_CHICK),
Xenopus: A4IID0 (A4IID0_XENTR), OMIM (human): 156845

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 73-108 Anti-MITF antibody, rabbit polyclonal, ChIP gradeFig.1 Western blot of Mitf in human melanoma cells.
Untreared:whole cell lysate
Control: Lysate of cells treated with control siRNA.
Mitf: Lysate of cells treated with siRNA against Mitf mRNA.
Anti-Mitf antibody was diluted at 1/1,000 in Can-Get-Signal Immunoreaction Enhanccer Solution(Toyobo,Tokyo). Upper band of Mitf is phosphorylated form of Mitf-M. The lower band is Mitf-M
isoform.
Anti-βactin antibody was as a loading control.

Fig.2 Immunofluorescence staining of melanocytes.
Mouse primary melanocytes 6 weeks after explantation was processed for immunofluorescencemicroscopy using anti-MiTF antibody at 1/500 dilution and DAPI

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Fig.3 Immunofluoresce staining of mouse melanoma cell.
Mouse melanoma B 16 cells are fixed with 4% PFA, permeabilized with 0.25 % Triton X-100.Anti-Mitf
antibody was used at 1/1,000 dilution.and as the secondary antibody, goat anti-rabbit IgG antibody(Alexa Fluor 488 conjugted) was used at 1/1,000 dilution.
Left: Immunofluoresencce staining with the antibody
Right: Merged image with nucleus stained with DAPI.

Fig.4 Immunohistochemical staining of Mitf in chick embrio at stage 14.
Embrio was fixed with paraformaldehyde and embedded in OCT compound and sectioned with acryostat at 8 μm. Anti-MiTF antibody was used at 1/300 dilution. As second antibody, Alexa 488
conjugated anti-rabbit IgG was used.
At stage 14, Mitif protein is detected throughout the RPE (Retinal Pigment Epithelium).
References: This antibody has been used in the following publications.
1. Osawa M et al. Molecular characterization of melanocyte stem cells in their niche. Development
132: 5589-5599 (2005) PMID: 16314490 IHC-Fr (ms)2. Delmas V et al.. β-Catenin induces immortalization of melanocytes by suppressing p16INK4aexpression and co-operates with N-Ras in melanoma development. Genes Dev 21: 2923-2935 (2007)
PMID: 18006687 IF (ms)
3. Yonetani S. et al. In Vitro Expansion of Immature Melanoblasts and their Ability to Repopulate
Melanocyte Stem Cells in the Hair Follicle. J Invest Dermatol. 2008 Feb;128(2):408-20.
PMID:17657242. IHC-Fr (ms)
4. Inoue-Narita T. et al. Pten deficiency in melanocytes results in resistance to hairgraying and susceptibility to carcinogen-induced melanomagenesis. Cancer Res. 2008
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Jul 15;68(14):5760-8. PMID:18632629 WB (ms)
5. Tsukiji N et al. Mitf functions as an in ovo regulator for cell differentiation and proliferation during
development of the chick RPE.Dev Biol 326: 335-346 (2009) PMID: 19100253

IHC-Fr, ChIP,(chicken)
6. Ohba K. et al. Microphthalmia-associated transcription factor is expressed inprojection neurons of the mouse olfactory bulb.Genes Cells. 2015 Dec;20 (12):1088-102. PMID:26522736. WB, IHC-P (ms)
7. Takeda K. et al. Regional Fluctuation in the Functional Consequence of LINE-1 Insertion in theMitf Gene: The Black Spotting Phenotype Arisen from the Mitfmi-bw Mouse Lacking Melanocytes.
PLoS One. 2016 Mar 1;11(3): PMID:26930598. IHC-P (ms)
8. Ohba K. et al. Microphthalmia-associated transcription factor ensures the elongation of axons anddendrites in the mouse frontal cortex. Genes Cells. 2016 Dec;21(12):1365-1379. PMID:27859996
IHC-P (ms)Related Products
73-107 Anti-MITF antibody, rabbit serum

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BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/