Rat mixed neuron/glial cultures stained with Myelin Basic Protein antibody 7D2 (red), and also with chicken antibody to neurofilament NF-L CPCA-NF-L
(green). Blue is a DNA stain. Note that the MBP 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 for MBP, though the neuronal processes can be seen with the NF-L antibody.
|Blots of 20µg crude rat brain homogenate were probed with 2 MBP antibodies; MCA-7G7 (lane 1) at 1:5,000 and MCA-7D2 (lane 2) at 1:5,000 dilution. MCA-7D2 binds the largest 21.5 kDa and 18.5 kDa transcripts preferentially, while monoclonal MCA-7G7 bind all four transcripts: 21.5kDa,18.5kDa, 17kDa and 14kDa.
||Mouse Monoclonal to Anti-Myelin Basic Protein, MBP
||18.5, and 21.5 kDa bands in rodents
||Purified myelin basic protein isolated from cow nerve.
||Antibody is supplied as an aliquot of ascites fluid
||Human, horse, cow, pig, chicken, rat, mouse
||Western blot, IF/ICC, IHC
|Suggestions for use
||Western blot: 1:5,000-1:10,000. IF/ICC and IHC: 1:1,000.
||Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles.
Myelin Basic Protein (MBP) is 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 mammalian 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 mammalian gene has 7 exons, and in humans exons 2 and 5 are either included or not, producing these four proteins. The single gene of rodents also produces 4 different proteins, but of slightly different sizes, 21.5, 18.5, 17 and 14 kDa. In this case the homologous exons are utilized differently: exons 2 and 6 are either inserted or not. 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 with a different monoclonal antibody our MCA-7G7. As shown below, MCA-7D2 only binds the two higher molecular weight isoforms of 21.5 kDa and 18.5 kDa in the rat. This antibody is therefore dependent on protein sequence encoded by exon 2, specifically the peptide KVPWLKPGRSPLPSHARSQPGLCNMY. The HGNC name for this protein is MBP.
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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