Exalpha Biologicals, Inc.

Product Highlight

FIX&PERM Cell Fixation and Permeabilization Kit

Flow cytometric analyses with monoclonal antibodies were so far mainly restricted to cell surface molecules. Intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins etc. were largely excluded from such studies. Also excluded from flow cytometric studies were cytoplasmic localizations of well-established membrane molecules like CD3 and CD22, which, in their cytoplasmic form, are the most reliable lineage markers in undifferentiated leukemia. With the FIX&PERM® Kit flow cytometric analysis of intracellular antigens has become as easy as surface antigen studies. The only prerequisite is the availability of suitable antibody conjugates. Most of the available monoclonal antibody conjugates can be used with the FIX&PERM® Kit, some determinants are sensitive, however, to the fixation step involved. This and the optimal fixation time have to be tested for each reagent.

News

Welcome, Peter Rutten

We are pleased to announce that Peter Rutten has now started in his role as Operations Director here at Exalpha Biologicals Inc. Peter has a Master’s of Science degree in Industrial and Organizational Psychology and this has lead him down a very business orientated career path. Peter is looking forward to working with the team at Exalpha. Peters primary focus is the customer experience and he will be working with the Laboratory team, the Quality Control team and the Order Processing team to ensure this focus is achieved. We all wish Peter well in his new role.

Exalpha Biologicals, Inc.

Microphthalmia Transcription Factor (MiTF)

  • Product Code: X1405M
  • Size: 100 µg
  • Availability: In Stock In Stock
  • Price (USD): $438

Cat #

X1405M		 Quantity:      

Data Sheet

Product Name

Microphthalmia Transcription Factor (MiTF)

Synonyms

MiTF

Host/Source

Mouse

Clone

C5

Isotype

IgG1

Product Type

Monoclonal Antibody

Reactivity

Human, Mouse, Rat

Applications

Gel Supershift, Western Blot, Immunoprecipitation, Immunohistochemistry (Frozen & Paraffin Sections)

Purification

Protein A/G Chromatography

Size

100 µg

Price (USD)

$438

Background

In Western blotting, it recognizes a doublet of 52-56kDa, identified as serine-phosphorylated and unphosphorylated forms of melanocytic isoforms of microphthalmia (Mi) transcription factor. 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 52kDa and 56kDa, while the longer Mi form runs as a cluster of bands at 60-70kDa in osteoclasts and in B16 melonoma cells (but not other melanoma cell lines), as well as mast cells and heart. It reacts with both melanocytic as well as the non- melanocytic isoforms of MiTF. This Ab does not cross-react with other b-HLH-ZIP factors by DNA mobility shift assay. Mi is a basic helix-loop-helix-leucin zipper (b-HLH-ZIP) transtripotion factor implicated in pigmentation, mast cells and bone development. The mutation of MiTF 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 MiTF including most of the b-HLH-ZIP region. Osteoclasts from these animals lack MiTF protein in contrast to wild-type rat, mouse, and human osteoclasts.

Immunogen

Hybridoma produced by the fusion of splenocytes from RBF/DnJ mice immunized with an N-terminal fragment of human microphthalmia protein and mouse myeloma NS1 cells.

Positive Control

501 Mel human melanoma cells, wild-type human, rat, mouse osteoclast cells

Formulation

Provided as solution in phosphate buffered saline with 0.08% sodium azide

Customer Storage

Product should be stored at -20°C. Aliquot to avoid freeze/thaw cycles

Target Molecular Weight

52-56kDa doublet

Product Image

Image Legend

Immunohistochemical staining using microphthalmia antibody on formalin-fixed, paraffin embedded EWS/ATF1-induced tumor cells (scale bar 100 µm). (Image modified from original taken from product specific reference 2)

Database Links:

SwissProtO75030Human

References

1. Weilbaecher KN, et. al. Age-resolving osteopetrosis: a rat model implicating microphthalmia and the related transcription factor TFE3. J. Exp.Med. 1998, 187: 775-785
2. Hemesath P, et. al. MAP kinase links the transcription factor microphthalmia to c-Kit signalling in melanocytes. Nature. 1998, 391:298-301


Product Specific References


This product has been used in:

1. Tshori, S., et al, 'Transcription factor MITF regulates cardiac growth and hypertrophy' Journal of Clinical Investigation 2006, 116, , 2673-2681
2. Yamada, K., et al. ?EWS/ATF1 expression induces sarcomas from neural crest?derived cells in mice.? J. Clin. Invest., 123, 600-610 (2013)
3. Hagglund, A.-C., et al. ?Canonical Wnt/ -Catenin Signalling Is Essential for Optic Cup Formation.? PLoS One, 8, e81158 (2013)
4. Nakano, T., et al. ?Self-formation of optic cups and storable stratified neural retina from human ESCs.? Cell Stem Cell (2012) 10, 771-785