Mouse Monoclonal Antibody to MAP2A/B (MAP2) Cat# MCA-5H11

      Microtubules are 25nm diameter protein rods found in most kinds of eukaryotic cells and are associated with a family of proteins called microtubule associated proteins (MAPs). MAPs play a crucial role in the regulation of microtubule dynamics and interactions in vivo. MAP2 was discovered as a high molecular weight MAP with an SDS-PAGE molecular weight of about 280kDa (1-3). A single mammalian MAP2 gene may generate two high molecular weight proteins of ~280kDa named MAP2A and MAP2B and lower molecular weight forms usually named MAP2C and MAP2D which run on SDS-PAGE gels at 60-70kDa. The 60-70kDa forms are found in neurons early in development, but are later replaced by the higher molecular weight forms (2). The MAP2A and MAP2B forms include a long protein sequence which forms fine filamentous protrusions from the sides of brain microtubules, which is therefore referred to as the projection domain. The epitope for this antibody was mapped to the projection domain so the antibody is specific for MAP2A and MAP2B. This region is one of the prototypes for "intrinsically unstructured regions", a widespread type of protein conformation (4). MAP2 isoforms are expressed only in neurons, specifically in the perikarya and dendrites of these cells. Antibodies to MAP2 isotypes are therefore excellent markers of neuronal dendrites and are useful for identifying neurons in cell culture and sections (e.g. 5-9).

Chromogenic Immunostaining of a formalin fixed paraffin embedded human brain cortex section with mouse mAb to MAP2, MCA-5H11, 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. MCA-5H11 strongly labels neurons, neuronal projections, and synapses. This antibody performs well in testing with 4% PFA fixed or standard NBF fixed rat and human tissue but does not stain long term NBF fixed material effectively. Due to its strong staining profile and ability to stain human tissue, MCA-5H11 is our recommended clone for MAP2 immunostaining. Mouse select image for larger view.



Chromogenic immunostaining of a 4% PFA fixed paraffin embedded rat cerebellum section with mouse mAb to MAP2, MCA-5H11, 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. In the cerebellum, MCA-5H11 strongly labels neurons, neuronal projections, and synapses. This antibody performs well in testing with 4% PFA fixed or standard NBF fixed rat and human tissue but does not stain long term NBF fixed material effectively. Due to its strong staining profile and ability to stain human tissue, MCA-5H11 is our recommended clone for MAP2 immunostaining. Mouse select image for larger view.



Immunofluorescent analysis of a rat cerebellum section stained with mouse mAb to MAP2, MCA-5H11, dilution 1:5,000 in green, and costained with rabbit pAb to α-internexin, RPCA-a-Int, dilution 1:2,000 in red. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 μM, and free-floating sections were stained with above antibodies. The MCA-5H11 antibody labels MAP2 protein in the perikarya and dendrites of the most neurons, particularly in granular cell and molecular layers of the cerebellum, while the α-internexin antibody selectively stains axons and dendrites of neuronal cells. Mouse select image for larger view.

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