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- add the lyophilized material (approx. 0.5 mg protein, mostly BSA) to 100 µl PBS or another buffered saline (add in order indicated) 

- allow some time for the material to dissolve; mix by swirling gently - remove particulate matter by centrifugation in a microfuge 

- add 100 µl glycerol (final glycerol conc. 50% v/v) and mix well (vortex) Final BSA-concentration in the stock solution: approx. 2.5 mg/ml.

Appropriate final dilutions should be determined experimentally. They may be in the range of 1:10,000 to 1:20,000 or even higher for floating sections, and around 1:1,000 or higher for mounted semithin sections (postembedding staining). These values are for combination with the PAP procedure. If the DAB reaction product in the PAP procedure is silver enhanced (see: Liu et al., Histochem., 1989, 90: 427-445), dilutions in the range of 1:5,000 to 1:20,000 or higher should be adequate for postembedding staining of semithin sections. Add 10% bovine serum to the diluted antibody

Store stock-solution in freezer (-20°C, not -70°C). Mix stock solution well before making aliquots. 10x diluted solutions may be kept in the refrigerator for some time (several weeks) but should be made 10 mM with sodium azide. Volumes of working dilutions (high dilutions) should be calculated such they are used up the same day. Avoid polystyrene tubes

1. Matute C, Streit P, Monoclonal antibodies demonstrating GABA-like immunoreactivity. Histochemistry 1986;86(2):147-57 2. Liu CJ, Grandes P, Matute C, Cuenod M, Streit P: Glutamate-like immunoreactivity revealed in rat olfactory bulb, hippocampus and cerebellum by monoclonal antibody and sensitive staining method. Histochemistry 1989;90(6):427-45