Parvalbumin Antibody

The second image above labeled "D." is parvalbumin (RED) interneurons pictured in mouse striatum, GREEN is GFP expressed by medium spiny neurons. Parvalbumin was used at 1:1000. This antibody performed very well. Scale bar = 25 microns. Immunohistochemistry: Mice were euthanized and perfused with PFA (4% in PBS), post fixed for 24 hrs, and their brains cryoprotected in 15% sucrose (24 hrs) followed by 30% sucrose (48 hrs). Coronal sections that included the corticostriatal, globus pallidus, or substantia nigra regions (30 um) were prepared using a microtome and then stored in PBS at 4C until processing. Sections were processed/stained in parallel as follows: Free floating sections were rinsed 3x with PBS and incubated 90 min at room temperature (RT) in PBS supplemented with donkey serum (5%) and Triton X-100 (1%). Primary antibody combinations were prepared as a master stock in PBS supplemented with donkey serum (2.5%) and Triton X-100 (0.5%) and applied to sections for 72 hrs at 4oC with gentle agitation. Sections were then rinsed 8x with PBS supplemented with Tween-20 (0.05%, at RT) and incubated with a secondary antibodies diluted in PBS supplemented with donkey serum (2.5%) and Triton X-100 (0.5%) for 1 hr at RT with gentle agitation, followed by 7 rinses with PBS and one rinse with deionized water. Sections were mounted onto slides and allowed to dry for ~18 hrs, after which cover slips were mounted with Vectashield and sealed with nail polish.