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 four different forms of the protein made by alternate transcription from a single gene, which have molecular weights of 21.5, 20.5, 18.5, and 17.2 kDa in humans. The single gene of rodents also produces 4 different proteins, but of slightly different sizes, 21.5, 18.5, 17 and 14 kDa. Sequence alignments of both sets of isotypes can be downloaded from here. We characterized our antibodies on rat spinal cord homogenates and detected the four bands expected for that species. Since this antibody binds all four bands it is therefore a pan-specific MBP antibody, recognizing a common epitope in all four gene products. The HGNC name for this protein is MBP.

Figures: Left: Blots of crude rat spinal cord homogenate blotted with three MBP antibodies; CPCA-MBP (first two lanes) MCA-7D2 (indicated lane) and MCA-7G7 (also as indicated). The MCA-7D2 monoclonal binds the largest 21.5kDa and 18.5kDa transcripts preferentially, while MCA 7G7 and CPCA-MBP bind all four transcripts. Right shows rat mixed neuron/glial cultures stained with Myelin Basic Protein monoclonal antibody MCA-7G7 (red). 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.

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 set of bands between of 21.5kDa and 14kDa for rodent and 21.5kDa to 17kDa for human.

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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).

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