General Lab Recipes
Lower Gel Buffer (4x Laemmli)
181.7 g Tris
4.0 g SDS
pH to 8.8 with HCl
q.s. with distilled water to 1 liter
Upper Gel Buffer (4x Laemmli)
60.6 g Tris
4.0 g SDS
pH to 6.8 with HCl
q.s. with distilled water to 1 liter
25T/1C Acrylamide [Neurotoxin - make in hood wearing gloves, goggles, and mask]
247 g Acrykamide
2.5 g Bis-Acrylamide
q.s. to 1 liter with distilled water
expansion upon stirring, don't add water all at once
filter through Whatman #1 paper
30T/2.7C Acrylamide [Neurotoxin - make in hood wearing gloves, goggles, and mask]
97.3 g Acrykamide
2.7 g Bis-Acrylamide
q.s. to 333 ml with distilled water
expansion upon stirring, don't add water all at once
filter through Whatman #1 paper
Running Buffer
30.3 g Tris base
144.1 g Glycine
10 g SDS
q.s. to 10 liters with distilled water
pH to 8.1 - 8.3 with con HCl if needed (should not need much if any pH'ing)
Gel Stain (Coomassie Blue)
1.0 g Coomassie Brilliant Blue
250 ml Methanol
75 ml Glacial Acetic Acid
q.s. to 1 liter with distilled water
Gel DeStain
75 ml Methanol
75 ml Glacial Acetic Acid
q.s. to 1 liter with distilled water
5x Sample Buffer (Stock)
15 ml Glycerol
15 ml 0.5M Tris HCl pH 6.8
10 ml 20% SDS (w/v)
15 mg Bromophenol Blue
q.s. to 100 ml with distilled water
[Bio Rad Mixture has 12.5 ml Tris, 10 ml Glycerol, and 10 ml SDS]
Sample Buffer (Working)
8 ml Stock Sample Buffer
1 ml 20% SDS
1 ml - mercaptoethanol (-ME)
mix well before use, use at least 20% sample buffer with sample, boil for > 4 min before loading on gel
Ammonium Persulfate
1 g Ammonium Persulfate
dissolve in 10 ml distilled water, aliquot into 1 ml microtubes and freeze at -20°C freezer