The buffer was prepared for one IPG strip of 7 cm, 50 μL of buffer including 12 mg (8 M) urea, 0.075 mg thiourea, 2 mg 4% CHAPS, 2.5 μL 100% (1 M) DTT, 0.2 μL 1% IPG buffer (ampholines), 20 μL Bromophenol blue, and dionized water up to 125 μL.
Standard buffers were prepared for both running (10X; Tris-Cl (30 g); Glycine (144 g); SDS (10 g)) and transfer buffer (10x; Tris-Cl (30 g); Glycine (144 g)).
Total lysates using radioimmunoprecipitation assay (RIPA) buffer were prepared and immunoblotted for protein expression studies using specific primary antibodies and appropriate secondary antibodies.
Samples consisting of serial dilutions of BoNT/A holotoxin in phosphate gelatin buffer were prepared, blinded, and shared with Sandia National Laboratories for parallel analysis via SpinDx, ELISA, and the live mouse bioassay on the same day.
Immunocomplexes were washed four times with NP-40 buffer containing 500 mM NaCl or RIPA buffer, and were prepared for Western blot analysis by boiling in 2× Laemmli loading buffer prior to electrophoresis on an 8% polyacrylamide gel.
Sections were then fixed in 1% osmium tetroxide/0.05 M phosphate buffer and were prepared for electron microscopic analysis.
To demonstrate growth in the media, and any effect of MES buffer, growth curves were prepared for the FN, the three NG, and the three LB (for the latter, from both low and high inocula), with and without MES, using the median bacterial count at each time point.
DAB working solution made up of 1 20 DAB chromogen in DAB substrate buffer was prepared and applied for 5 min.
After removal, the colons were fixed in 10% buffered formalin for immunohistochemistry, protein lysates were prepared for western analysis and ELISA.
Samples were prepared for ESR by buffer-exchanging them into H2O/deuterated ethylene-glycol medium [Tes (pH 7.4) and 100 mM NaCl].
