The Immunogen: Myelin Basic Protein (MBP) s one of the major proteins of the myelin sheath surrounding axons in the nervous system. Since it is of relatively low molecular weight and high abundance the protein sequence was determined from purified protein over 30 years ago (1). The protein is made by oligodendrocytes in the central nervous system, so antibodies to MBP are good markers of this cell type. However, transcripts from the same gene are also expressed in certain hematopoetic lineage cells (2). In the central nervous system there are there different forms of the protein made by alternate transcription from a single gene, which have molecular weights of 21.5, 18.5, and 17.2 kDa. Since the two lower molecular weight forms are very close in molecular size, MBP antibodies typical show two bands on Western blots, one at about 22kDa and another at about 18kDa.

Antibody Characteristics: Antibody was raised in chicken against petide sequences conserved in higher vertebrate MBP proteins. There different peptide antibodies were used, each was affinity purified on the appropriate peptide, and then mixed to produce this antibody preparation. This antibody is an IgY preparation, with total protein content about 200 microgram/ml. The preparation contains 10mM sodium azide as a preservative. Store at 4°C or -20°C. Avoid repeat freezing and thawing.

Figures: Left: Blots of whole tissue homogenate of rat spinal cord with chicken antibody to Myelin Basic Protein. The antibody stains a prominent ~20kDa band at the dye front on this 12% acrylamide gel. Right shows rat mixed neuron/glial cultures stained with chicken Myelin Basic Protein (green). Blue is a DNA stain. Note that the Myelin Basic Protein antibody stains an oligodendrocyte and some membrane shed from this cell. Other cells in the field include neurons, astrocytes, microglia and fibroblasts, all of which are completely negative. specifically, and that the staining is concentrated in the cell bodies,

Suggestions for use: Try at dilutions of 1:1,000 and higher for immunofluorescence. For western blots try at 1:10,000. A suitable control tissue is rat spinal cord or peripheral nerve homogenate. The major isoforms of MBP run as a closely spaced double of 22kDa and 18kDa.

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References:

1. Eylar EH, Brostoff S, Hashim G, Caccam J, Burnett P. Basic A1 protein of the myelin membrane. The complete amino acid sequence. J. Biol. Chem. 246:5770-5784 (1971).

2. Marty MC, Alliot F, Rutin J, Fritz R, Trisler D. and Pessac B. The myelin basic protein gene is expressed in differentiated blood cell lineages and in hemopoietic progenitors. Proc. Nat. Acad. Sci. 99:8856-8861 (2002).

Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.

Availability and Price: Available for shipping now, $200 US per aliquot of 500 microliters of IgY preparation, enough for hundreds of experiments. For order form press here

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