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MAB3747-I Anti-Microphthalmia (Mi) Antibody, clone C5

MAB3747-I
100 µg  
Purchase on Sigma-Aldrich

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Overview

Replacement Information

Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
H, MWB, IHC, IF, ICC, EMSA, IPMPurifiedMonoclonal Antibody
Description
Catalogue NumberMAB3747-I
DescriptionAnti-Microphthalmia (Mi) Antibody, clone C5
Alternate Names
  • Microphthalmia-associated transcription factor
  • Class E basic helix-loop-helix protein 32
  • bHLHe32
Background InformationMiTF (Microphthalmia associated transcription factor) is a basic helix loop helix leucine zipper (b HLH ZIP) transcription factor implicated in pigmentation, mast cells and bone development. Mutations in MiTF cause auditory pigmentary syndromes, such as Waardenburg syndrome type II, type IIa and Tietz syndrome in humans. 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 52 kDa and 56 kDa, while the longer Mi form runs as a cluster of bands at 60-70 kDa in osteoclasts and in B16 melonoma cells (but not other melanoma cell lines), as well as mast cells and heart. MiTF plays a critical role in the differentiation of various cell types as neural crest-derived melanocytes, mast cells, osteoclasts and optic cup-derived retinal pigment epithelium. Mi is a basic helix-loop-helix-leucine zipper (b-HLH-ZIP) transcription factor implicated in pigmentation, mast cells and bone development. The mutation of Mi 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 Mi including most of the b-HLH-ZIP region. Osteoclasts from these animals lack Mi protein in contrast to wild-type rat, mouse, and human osteoclasts.
References
Product Information
FormatPurified
Control
  • Mouse brain tissue lysates
PresentationPurified mouse monoclonal IgG2aκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationUse Anti-Microphthalmia (Mi), clone C5 mouse monoclonal antibody validated in Electrophoretic Mobility Shift Assay (EMSA), Immunocytochemistry, Immunohistochemistry, Immunoprecipitation and Western blotting for the detection of Microphthalmia-associated transcription factor.
Key Applications
  • Western Blotting
  • Immunohistochemistry
  • Immunofluorescence
  • Immunocytochemistry
  • Electrophoretic Mobility Shift Assay
  • Immunoprecipitation
Application NotesImmunohistochemistry Analysis: A representative lot detected microphthalmia immunoreactivity in formalin-fixed, paraffin-embedded human metastatic melanoma tissue sections by fluorescent immunohistochemistry (Feige, E., et. al. (2011). Proc. Natl .Acad. Sci. U. S. A. 108(43):E924-E933).
Immunocytochemistry Analysis: A representative lot detected the exogenously expressed murine microphthalmia mutant constructs, Mitf D222/236N and Mitf D222N (mi-vit), in the nucleus of transfected COS-7 cells. Dual staining showed much reduced β-catenin-anchoring ability of these mutants in the nucleus (Schepsky, A., et al. (2006). Mol. Cell. Biol. 26(23): 8914-8927).
Immunocytochemistry Analysis: A representative lot detected a time-dependent induction of microphthalmia upregulation in B16/F10 murine melanoma cells upon Forskolin stimulation by fluorescent immunocytochemistry (Bertolotto, C., et al. (1998). J. Cell Biol. 142(3):827-835).
Electrophoretic Mobility Shift Assay (EMSA): A representative lot caused a supershift of Mbox motif oligonucleotide-complexed wild-type and D222/236N and D222N mutant murine microphthalmia constructs by EMSA (Schepsky, A., et al. (2006). Mol. Cell. Biol. 26(23): 8914-8927).
Electrophoretic Mobility Shift Assay (EMSA): A representative lot caused a supershift of Mbox motif oligonucleotide-complexed microphthalmia, but not TFE3-DNA complex by EMSA using in vitro translated microphthalmia and TFE3 or B16/F10 murine melanoma cell nuclear extract (Verastegui, C., et al. (2000). Mol. Endocrinol. 14(3):449-456).
Immunoprecipitation Analysis: A representative lot immunoprecipitated microphthalmia from B16/F10 murine melanoma cell nuclear extracts (Verastegui, C., et al. (2000). Mol. Endocrinol. 14(3):449-456).
Western Blotting Analysis: A representative lot detected microphthalmia expression in murine splenocytes and B16/F10 murine melanoma cells (Verastegui, C., et al. (2000). Mol. Endocrinol. 14(3):449-456).
Western Blotting Analysis: A representative lot detected a time-dependent induction of microphthalmia upregulation in B16/F10 murine melanoma cells and normal human melanocytes upon stimulation by Forskolin or α-melanocyte–stimulating hormone (αMSH) (Bertolotto, C., et al. (1998). J. Cell Biol. 142(3):827-835).
Biological Information
ImmunogenRecombinant N-terminal fragment of human microphthalmia protein.
EpitopeN-terminal
CloneC5
ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
HostMouse
SpecificityIn Western blotting, it recognizes a doublet of 52-56 kDa, identified as serine-phosphorylated and unphosphorylated forms of melanocytic isoforms of microphthalmia (Mi). There are two known isoforms of Mi differing by 66 amino acids at the NH2 terminus. Shorter forms are expressed in melanocytes and run as two bands at 52 kDa and 56 kDa, while the longer Mi form runs as a cluster of bands at 60-70 kDa in osteoclasts and in B16 melonoma cells (but not other melanoma cell lines), as well as mast cells andheart. It reacts with both melanocytic as well as the nonmelanocytic isoforms of Mi. This Ab does not cross-react with other b-HLH-ZIP factors by DNA mobility shift assay.
IsotypeIgG1κ
Species Reactivity
  • Human
  • Mouse
Species Reactivity NoteHuman and Mouse. Predicted to react with Rat based on sequence homology.
Antibody TypeMonoclonal Antibody
Entrez Gene Number
Gene Symbol
  • MITF
  • BHLHE32
  • MiTF
Purification MethodProtein G Purified
UniProt Number
Molecular Weight~52/56 kDa observed. An uncharacterized band appears at ~140 kDa in some lysates.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceEvaluated by Western Blotting in mouse brain tissue lysate.

Western Blotting Analysis: An 1:500 dilution of this antibody detected Microphthalmia in 10 µg of mouse brain tissue lysate.
Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
Packaging Information
Material Size100 µg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
MAB3747-I 04053252691942

Documentation

Anti-Microphthalmia (Mi) Antibody, clone C5 SDS

Title

Safety Data Sheet (SDS) 

Anti-Microphthalmia (Mi) Antibody, clone C5 Certificates of Analysis

TitleLot Number
Anti-Microphthalmia (Mi), clone C5 - 2360618 2360618
Anti-Microphthalmia (Mi), clone C5 - 2309258 2309258
Anti-Microphthalmia (Mi), clone C5 - 3193863 3193863
Anti-Microphthalmia (Mi), clone C5 - 3301919 3301919
Anti-Microphthalmia (Mi), clone C5 - 3731760 3731760
Anti-Microphthalmia (Mi), clone C5 - 3944861 3944861
Anti-Microphthalmia (Mi), clone C5 - 4019799 4019799
Anti-Microphthalmia (Mi), clone C5 - 4113823 4113823
Anti-Microphthalmia (Mi), clone C5 - Q2019899 Q2019899
Anti-Microphthalmia (Mi), clone C5 -2505485 2505485

References

Reference overviewPub Med ID
Melanogenesis-inducing effect of cirsimaritin through increases in microphthalmia-associated transcription factor and tyrosinase expression.
Kim, HJ; Kim, IS; Dong, Y; Lee, IS; Kim, JS; Kim, JS; Woo, JT; Cha, BY
International journal of molecular sciences  16  8772-88  2015

Show Abstract
25903150 25903150
Malignant PEComa of the adrenal gland.
Lau, Sean K
Pathol. Res. Pract., 208: 113-7 (2012)  2012

Show Abstract
22154607 22154607