Immunofluorescent analysis of adult rat hippocampus section stained with Anti-MAP2 Antibody [2C4] (A85459), at a dilution of 1:5,000 in green, and Anti-FOX2 Antibody (A104328), at a dilution of 1:2,000, in red. Following transcardial perfusion of the rat with 4% paraformaldehyde, the brain was post-fixed for 24 hours, cut to 45 µm, and free-floating sections were stained with the above antibodies. Anti-MAP2 Antibody [2C4] (A85459) labels all MAP2 protein isotypes expressed in neuronal perikarya and dendrites. The Anti-FOX2 Antibody (A104328) stains the nuclei of most neuronal cells.
Mixed neuron and glia cultures stained with Anti-MAP2 Antibody (red) and Anti-Vimentin Antibody (A85421 | green) and DNA (blue). Anti-MAP2 Antibody reveals strong cytoplasmic staining of dendrites and perikarya of neuronal cells, while vimentin was visualized in astrocytes and fibroblasts.
Figure 3: Western Blot - Anti-MAP2 Antibody [2C4] (A85459)
Western blot analysis of tissue and cell lysates using Anti-MAP2 Antibody [2C4] (A85459), at a dilution of 1:5,000, in green. The lanes contain samples of: [1] Protein standards, in red, [2] rat whole brain lysate, [3] HeLa cells, [4] SH-SY5Y cells, [5] HEK293 cells and [6] NIH-3T3 cell lysates. A band at about 280 kDa corresponds to the full length intact MAP2A/2B isotypes, while bands at about 70 kDa represent MAP2C/D isotypes. Multiple bands in between are likely in vivo fragments of MAP2A/B. Only the SH-SY5Y cells, which have neuronal properties, express MAP2 protein. The same blot was simultaneously probed with Anti-Vimentin Antibody (A85421), at a dilution of 1:5,000, in red. The Anti-Vimentin Antibody (A85421) binds to a single band at approximately 50 kDa present in all preparations, so acts as a positive loading control.
Figure 4: Western Blot - Anti-MAP2 Antibody [2C4] (A85459)
Western blot of Rat whole brain extract, Hela, SH-SY5Y, HEK293, and NIH/3T3 cells probed with Anti-MAP2 Antibody (green). All forms of MAP2 are present in the rat brain homogenate. A band at ~280 kDa corresponds to full length intact MAP2A/2B. Bands appearing at 70kDa correspond to MAP2C/D. The fainter bands in between correspond to in vivo fragments of MAP2A/B, which are very sensitive to proteases. Of the four cell lines tested, only the SH-SY5Y cell line, which has neuronal properties, is positive for ~70kDa MAP2C/D. Blot was simultaneously probed with Anti-Vimentin Antibody (A85421 | red), showing a single strong band at ~50 kDa.
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