Nuclear protein extracts from the cerebellar vermis, prefrontal cortex, amygdala, and hippocampus were solubilized in a sample buffer (0.125 M Tris base; 4% SDS; 20% v/v glycerol; 0.2 M DTT; 0.02% bromophenol blue
, pH 6.8) at a concentration of 10 mg/15 [micro]L per well and boiled for 5 min at 95[degrees]C.
The reaction was then stopped by addition of Tris-HCl buffer (125 mM), pH 6.8 containing 10% of glycerol, 5 of [beta]-mercaptoethanol, 2 of SDS and 0.001% of bromophenol blue
. Samples were boiled for 5 min.
Lugol's test was used for detection of starch (Johansen, 1940); bromophenol blue
for total protein (Baker, 1958); Sudan Red B for total lipids (Brundett, Kendrick, & Peterson, 1991); Fehling's reagent for reducing sugars (Sass, 1951); and the DAB test "0.5% 3,3'diaminobenzidine" (DAB Sigma[R]) for hydrogen peroxide (Rossetti & Bonnatti, 2001).
Sucrose 40% and bromophenol blue
(0.25%) were mixed with sterile double distilled water.
Five IPG Immobiline DryStrips 24 cm, pH 3 to 10 (GE Healthcare, Bucks, United Kingdom) were rehydrated overnight in 450 [micro]l 2DE rehydration buffer (7M urea, 2M thiourea, 2% CHAPS, 0.5% pH 3 to 11 ampholytes (GE Healthcare), 1% DTT, trace bromophenol blue
The protein samples were prepared in 1x SDS loading buffer (2% (w/v) SDS, 5% (v/ v) glycerol, 0.01% (w/v) bromophenol blue
, 8% (w/v) DTT, which was added just before heating) and heated at 100 [degrees]C for 5 min.
Aliquots of each sample were mixed with 2x sample dissolving buffer (4% SDS, 20% glycerol, 0.125 M Tris-HCl buffer pH 6.8, 0.02% bromophenol blue
The color change is due to the production of lactic acid, and the pH was reduced so the color of bromophenol blue
was changed from blue to yellow color.
Bahadur, "Enhancement of power conversion efficiency of dye-sensitized solar cells by co-sensitization of Phloxine B and Bromophenol blue
dyes on ZnO photoanode," Journal of Luminescence, vol.
They included bromocresol green, bromophenol blue
, bromothymol blue, bromocresol purple, bromocresol blue, cresol red, cresol blue, cresol purple (BDH Chemicals Ltd., Poole, England); Diaverdine (Elnaser pharmaceutical chemicals Cairo, Egypt); and New-Cox powerful anticoccidial watersoluble powder (Atco Pharma for Pharmaceutical Industry, Cairo, Egypt).
Acrylamide, bis-acrylamide, ammonium persulfate (APS), N,N,N,N-Tetramethylethylenediamine (TEMED), sodium dodecyl sulfate (SDS), tris(hydroxymethyl)-aminomethane, glycine, bromophenol blue
, [beta]-mercaptoethanol, and Coomassie Brilliant Blue G-250 were purchased from Bio-Rad Laboratories (Hercules, CA).