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Merck
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資料

安全性情報

MAB3747-I

Sigma-Aldrich

Anti-Microphthalmia (Mi) Antibody, clone C5

clone C5, from mouse

別名:

Microphthalmia-associated transcription factor, Class E basic helix-loop-helix protein 32, bHLHe32

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About This Item

UNSPSCコード:
12352203
eCl@ss:
32160702
NACRES:
NA.41

由来生物

mouse

品質水準

抗体製品の状態

purified immunoglobulin

抗体製品タイプ

primary antibodies

クローン

C5, monoclonal

化学種の反応性

mouse, human

化学種の反応性(ホモロジーによる予測)

rat

テクニック

electrophoretic mobility shift assay: suitable
immunocytochemistry: suitable
immunofluorescence: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

アイソタイプ

IgG2aκ

NCBIアクセッション番号

UniProtアクセッション番号

輸送温度

wet ice

ターゲットの翻訳後修飾

unmodified

遺伝子情報

human ... MITF(4286)

詳細

MiTF (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.

特異性

In 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.

免疫原

Recombinant N-terminal fragment of human microphthalmia protein.

アプリケーション

Immunohistochemistry 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).
Use 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.

品質

Evaluated 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.

ターゲットの説明

~52/56 kDa observed. An uncharacterized band appears at ~140 kDa in some lysates.

物理的形状

Format: Purified

アナリシスノート

Control
Mouse brain tissue lysates

その他情報

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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保管分類コード

12 - Non Combustible Liquids

WGK

WGK 1

引火点(°F)

Not applicable

引火点(℃)

Not applicable


適用法令

試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。

Jan Code

MAB3747-I:


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