| Name: | Mouse Monoclonal to Myelin Basic Protein, MBP |
| Immunogen: | Purified myelin basic protein isolated from bovine brain, epitope is in peptide TPPPSQGKG, amino acids 125-133 of the human 21.5kDa sequence |
| HGNC Name: | MBP |
| UniProt: | P02687 |
| Molecular Weight: | 14-21.5kDa rodent, 17.2-21.5kDa human |
| Host: | Mouse |
| Isotype: | IgG1 heavy, κ light |
| Species Cross-Reactivity: | Human, rat, mouse, cow, pig, horse |
| RRID: | AB_2572353 |
| Format: | Purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3 |
| Applications: | WB, IF/ICC, IHC |
| Recommended Dilutions: | WB: 1:10,000 -1:20,000. IF/ICC 1:2,000-5,000. IHC: 1:10,000. |
| Storage: | Store at 4°C for short term, for longer term at -20°C. |
Immunofluorescent analysis of rat hippocampal section stained with mouse mAb to myelin basic protein (MBP), MCA-7G7, dilution 1:5,000 in green, and costained with rabbit pAb to neurofilament NF-H, RPCA-NF-H, dilution 1:2,000, in red. The MBP antibody stains oligodendrocyte cell bodies and the myelin sheathes around axons, while the NF-H antibody labels the axons themselves.
Western blot analysis of different tissue lysates using mouse mAb to MBP, MCA-7G7, dilution 1:20,000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord, [4] rat sciatic nerve, [5] mouse brain, [6] mouse spinal cord. Multiple bands at 14kDa, 17kDa, 18.5kDa to 21.5kDa are the alternate transcripts of MBP. Other bands are proteolytic fragments of the MBP protein.
Mouse Monoclonal Antibody to Myelin Basic Protein
Cat# MCA-7G7
$120.00 – $800.00
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 and nervous system, so antibodies to MBP are good markers of this cell type. In the peripheral nervous system MBP is expressed by myelinating Schwann cells so this antibody can be used to identify these cells in culture or sectioned materials. In the central nervous system four different forms of the protein made by alternate transcription from a single gene, the protein products having molecular weights of 21.5, 20.5, 18.5, and 17.2kDa in humans. The single gene of rodents also produces 4 different proteins but the splicing mechanism is different producing four forms of slightly different sizes, 21.5, 18.5, 17 and 14kDa. Some interest has focused on MBP as a potentially significant auto-antigen involved in mouse models of multiple sclerosis (MS, 3) and in human patients (4). Detection of MBP released into blood and CSF has some potential as a surrogate biomarker of demyelination and axonal loss in MS and other relevant damage and disease states (e.g. 5).
The MCA-7G7 antibody was made against a preparation of MBP purified biochemically from bovine brain. It can be used to identify oligodendrocytes and Schwann cells in neural cell culture, to visualize myelin sheaths and myelinating cells in sectioned material and to probe western blots for MBP gene products. The antibody is also rather insensitive to aldehyde fixation and so can be used in immunohistochemistry of paraffin sections. The MCA-7G7 antibody binds all four of the CNS MBP isoforms, so that the epitope for the antibody is located in the “core” shared by all four gene products. Further mapping localizes the epitope to peptide TPPPSQGKG, amino acids 125-133 of the human 21.5kDa sequence. The data was produced with overlapping peptides which suggests that the last four amino acids, SQGKG, are likely to be key elements of the epitope. This peptide is invariant in rat, mouse, cow and many other species, so this antibody will have wide applicability. In contrast our alternate mouse monoclonal MCA-7D2 binds only the 21.5kDa and 18.5kDa rat MBP isotypes but all bovine and human isotypes, mapping the epitope to AEGQRPGFGYGGRASDYKSAHKGFKGVDAQGTLSKIFKLG, amino acids 145-184 of the human 21.5kDa sequence. A sequence alignment of the four CNS MBP isotypes in human and rat can be downloaded from here. The antibody works well for western blotting and for IF, ICC and IHC (for IHC see data under “Additional Info” tab). Mouse select image at left for larger view.
Chromogenic immunostaining of a 4% PFA fixed paraffin rat hippocampus section with mouse mAb to MBP, MCA-7G7, dilution 1:5,000, detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. The MBP antibody labels axonal myelin sheaths. This antibody performs well in testing with both 4% PFA and standard NBF fixed rat and human tissues. Mouse select image for larger view.
Above: Immunofluorescent analysis of rat brain stem section stained with mouse mAb to Myelin Basic Protein (MBP), MCA-7G7, dilution 1:5,000 in green, and costained with rabbit pAb to neurofilament NF-H, RPCA-NF-H, dilution 1:2,000 in red. The MBP antibody stains oligodendrocytes and the myelin sheathes around axons. In this high magnification view it is clear that the NF-H antibody labels axons within the myelin sheathes. Mouse select image for larger view.
Above: Blots of 20µg crude rat brain homogenate blotted with 2 MBP antibodies; MCA-7G7 (left lane) at 1:5,000 and MCA-7D2 (right lane) also at 1:5,000 dilution. MCA-7G7 bind all four rat transcripts running at 21.5kDa, 18.5kDa, 17kDa and 14kDa, while the alternate EnCor antibody MCA-7D2 binds only the two largest transcripts. We can conclude MCA-7G7 binds the core region of MBP while MCA-7D2 binds to the sequence encoded by exon 2 of the MBP gene. A sequence alignment of the human and rat MBP alternate transcripts is here.
Above: 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 with this antibody. Mouse select image for larger view.
A sequence alignment of the four CNS MBP isotypes in human and rat can be downloaded here.
1. Eylar EH, et al. Basic A1 protein of the myelin membrane. The complete amino acid sequence. J. Biol. Chem. 246:5770-84 (1971).
2. Marty MC, et al. The myelin basic protein gene is expressed in differentiated blood cell lineages and in hemopoietic progenitors. PNAS 99:8856-61 (2002).
3. Libbey JE, Fujinami RS. Experimental Autoimmune Encephalomyelitis as a Testing Paradigm for Adjuvants and Vaccines. Vaccine 29:3356–62 (2011).
4. Wucherpfennig KW, Strominger JL. Molecular mimicry in T cell-mediated autoimmunity: Viral peptides activate human T cell clones specific for myelin basic protein. Send to
Cell 80:695-705 (1995).
5. Berger RP, et al. Serum neuron-specific enolase, S100B, and myelin basic protein concentrations after inflicted and noninflicted traumatic brain injury in children. J. Neurosurg. 103:61-8 (2005).
