Immunofluorescent analysis of rat cerebellum section stained with Anti-Calretinin Antibody (1:500 | green) and Anti-Calbindin Antibody (A85359 | 1:2,000 | red). Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45µM, and free-floating sections were stained with the above antibodies. The Anti-Calretinin Antibody labels interneurons in the granule cell layer, while the Anti-Calbindin Antibody stains Purkinje neurons and their dendritic processes in the molecular layer of the cerebellum.
Left: Adult mouse brain hippocampal section (45 µM; fixed by transcardial perfusion with 4% paraformaldehyde) was co-stained with Anti-Calretinin Antibody (1:1000 | red) and Anti-Fox3/NeuN Antibody (A85403 | 1:1,000 | green). Anti-Calretinin Antibody stains a small number of interneuron in the stratum radiatum of CA1 region, and Fox/NeuN is expressed in most neurons in the brain. As a result, cells are positive for calretinin appear to be yellow. Right: Adult rat cortical section (45 µM; fixed by transcardial perfusion with 4% paraformaldehyde) was co-stained with Anti-Calretinin Antibody (red) and Anti-Calbindin Antibody (A85359 | green). In cortex, calretinin is expressed in a small population of interneurons concentrated in Layer 4 area, while calbindin is expressed in cells concentrated in Layer 2/3. Because each antibody specifically labels a different population of cells exclusively, the cells are either stained with green or red in the cortex. Insets are high-magnification images of the boxed area in each picture. Blue is DAPI staining that labels DNA.
Figure 3: Western Blot - Anti-Calretinin Antibody [3G9] (A85367)
Western blot analysis of recombinant protein solutions using Anti-Calretinin Antibody [3G9] (A85367), at a dilution of 1:1,000, in green. The lanes contain samples of: [1] Protein standards, in red, [2] calbindin, [3] calretinin, [4] parvalbumin, and [5] secretogogin. The Anti-Calretinin Antibody [3G9] (A85367) reacts only with calretinin, and does not bind to other related calcium-binding proteins.
Figure 4: Western Blot - Anti-Calretinin Antibody [3G9] (A85367)
Western blot analysis of Anti-Calretinin Antibody. Left: Blot of cow cerebellum lysate was probed with Anti-Calretinin Antibody. The antibody binds strongly and cleanly to the calretinin band at 29kDa. Right: Blot of recombinant protein: secretagogin (Lane 1), parvalbumin (Lane 2), calretinin (Lane 3), calbindin (Lane 4) was probed with Anti-Carletinin Antibody. Lane 5 is protein marker. This antibody recognizes only calretinin, not other calcium-binding proteins.
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