Stock buffer / solutions list

Acrylamide for separating gel (Acrylamide : BIS = 30 : 0.135) Acrylamide 30.00 g ———————————————————————————————————————————————————— BIS 0.135 g Make volume to 100 ml with MQ water. Keep in dark (Brown bottle) Separating gel buffer (pH 8.8) (Final Conc.) Tris 12.11 g 1 M ———————————————————————————————————————————————————— SDS 0.27 g 0.27% Dissolve in 80 ml MQ water, adjust pH to 8.8, make the vol. to 100 ml Acrylamide for stacking gel (Acrylamide : BIS = 29.2 : 0.8) Acrylamide 29.2 g ———————————————————————————————————————————————————— BIS 0.8 g Make volume to 100 ml with MQ water. Keep in dark (Brown bottle) Stacking gel buffer (pH 6.8) (Final Conc.) Tris 3.03 0.25 M ———————————————————————————————————————————————————— SDS 0.20 g 0.2% Dissolve in 80 ml MQ water, adjust pH to 6.8, make the vol. to 100 ml SDS-PAGE running buffer Tris 9.0 g Glycine 43.2 g ———————————————————————————————————————————————————— SDS 3.0 g Dissolve in 3 L MQ water SDS-sample buffer Glycerol 10 ml Tris 0.757 g SDS 2.5 g ———————————————————————————————————————————————————— 2-Mercaptoethanol 5.0 ml Dissolve in 100 ml MQ water Bromophenol blue (BPB) solution Dissolve 0.1 g BPB in 100 ml 10% glycerol Blotting solution A (Final conc.) Tris 36.33 g 0.3 M Methanol 200 ml 20% ———————————————————————————————————————————————————— SDS 0.20 g 0.02% Make vol to 1000 ml with MQ water, keep at 4 ºC Blotting solution B (Final conc.) Tris 3.03 g 25 mM Methanol 200 ml 20% SDS 0.20 g 0.02% ———————————————————————————————————————————————————— Make vol to 1000 ml with MQ water, keep at 4 ºC Blotting solution C (Final conc.) Tris 3.03 g 25 mM s- Amino-n-Caproic Acid 5.20 g 40 mM Methanol 200 ml 20% SDS 0.20 g 0.02% ———————————————————————————————————————————————————— Make vol to 1000 ml with MQ water, keep at 4 ºC Ponceau 3S staining solution Dissolve 0.1 g Ponceau 3S in 100 ml 1% Acetic acid Coomassie brilliant blue (CBB) solution Methanol 1500 ml Acetic acid 300 ml Coomassie brilliant blue-R250 3 g ———————————————————————————————————————————————————— Dissolve in 3000 ml MQ water Destaining solution Methanol 1100 ml Acetic acid 300 ml ———————————————————————————————————————————————————— Dissolve in 3000 ml MQ water Acrylamide solution for 1D-PAGE(Acrylamide : BIS = 28.38 : 1.62) Acrylamide(High grade) 28.38 g BIS 1.62 g ———————————————————————————————————————————————————— Make vol to 1000 ml with MQ water, keep in dark Lysis buffer Urea 4.8 g NP - 40 0.2 ml Ampholine(pH 3.5-10.0) 0.2 ml 2-Mercaptoethanol 0.5 ml Polyvinyl pyrrolidone-40 0.5 g ———————————————————————————————————————————————————— Make vol to 10 ml with MQ water, presurve in 2 ml eppendorf tube at -20 4 ºC 0.02 N H3PO4 Add 0.48 ml H3PO4 (42 N, specific weight 1.87, 72%) to 1000 ml MQ water 0.02 N NaOH Dissolve 0.8 g NaOH to 1000 ml MQ water.