Anti-Calretinin Antibody [6A9] (A85366)

Immunofluorescent analysis of a section of rat hippocampus section stained with Anti-Calretinin Antibody, at a dilution of 1:2,000, in green, and co-stained with Anti-Parvalbumin Antibody (A85316 | 1:1,000, in red. The blue is DAPI staining of nuclear DNA. 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 above antibodies. The calretinin and parvalbumin antibodies stain different subsets of GABAergic interneurons.

Left: Mixed neuron/glial cultures stained with Anti-Calretinin Antibody, at a dilution of 1:2,000, in red, and Anti-Vimentin Antibody (A85421 | 1:5,000, in green. Calretinin is prominently expressed in small number of interneurons, while astrocytes and fibroblasts were visualized with the Anti-Vimentin Antibody. Middle: Adult rat cortical section (45 µM; fixed by transcardial perfusion with 4% paraformaldehyde) was co-stained with Anti-Calretinin Antibody, at a dilution of 1:1,000, in red, and Anti-Calbinidin Antibody (A85365 | 1:1,000, in green. In the motor cortex, calretinin is expressed in a small population of interneurons that do not express calbindin. Because each antibody specifically labels a different population of cells exclusively, the cells are either stained with red or green. Right: Adult mouse brain hippocampal section (45 µM; fixed by transcardial perfusion with 4% paraformaldehyde) was co-stained with Anti-Calretinin Antibody, at a dilution of 1:1,000, in red, and Anti-Calbindin Antibody (A85365 | 1:1,000, in green. In the stratum radiatum of CA1 region, calretinin expresses in a small number of interneurons that do not express calbindin. As a result, our antibodies label different neurons in either red or green. Insets are high-magnification images of the boxed area in each picture. Blue is a hoechst DNA staining.

Western blot analysis of tissue lysates probed with Anti-Calretinin Antibody, at a dilution of 1:2,000, in red,: [Lane 1] protein standard (red), [Lane 2] rat brain, [Lane 3] rat spinal cord, [Lane 4] mouse brain, [Lane 5] mouse spinal cord, and [Lane 6] cow spinal cord. The single clean band at 29kDa corresponds to the calretinin protein.

Western blot analysis of Anti-Calretinin Antibody. Blot of rat brain lysates (Lane 1), recombinant proteins: pavalbumin (Lane 2), calretinin (Lane 3), calbindin (Lane 4) was probed with Anti-Calretinin Antibody, at a dilution of 1:5,000). In rat brain lysates, this antibody recognises a clean band at 29 kDa which represents calretinin. Also it reacts with only calretinin, not other calcium-binding proteins.