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 including MCA-7G7 typical show two bands on Western blots, one at about ~22kDa and another at about ~18kDa. The HGNC name for this protein is MBP.

Figures: Left: Blots of crude rat spinal cord homogenate blotted with MCA-7D2 (right lane) and MCA-7G7 (left lane). The MCA-7D2 monoclonal binds only the largest 21.5kDa transcript, while MCA 7G7 binds all three 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 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).

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