SDS-PAGE Reagents
U.K. Laemmli. 1970. "Cleavage of Structural Proteins During the Assembly of the Head of Bacteriophage T4." Nature 227: 680-685. (The founding SDS-PAGE reference.)

Stock Gel Solution

"A/B" = 29.2% acrylamide; 0.8% bis-acrylamide
(these are carcinogens but reputedly less so than a single cigarette)
q.s. 100 ml with glass distilled water (yields world-class gels!)
NOT deionized water - this is by far the worst!
NOT metal distilled water
Tap water works better than deionized water;
Seawater works even better than deionized water!
(Reasons: polyvalent cations and dissolved ion-exchange resins bind to macromolecules and distort their electrophoretic mobilities.)
(A/B good stored at RT for weeks; good for years at 4°c)

"A/B"	= 29.2% acrylamide; 0.8% bis-acrylamide (these are carcinogens but reputedly less so than a single cigarette)
q.s. 100 ml with glass distilled water (yields world-class gels!)
NOT deionized water - this is by far the worst!
NOT metal distilled water
Tap water works better than deionized water;
Seawater works even better than deionized water!
(Reasons: polyvalent cations and dissolved ion-exchange resins bind to macromolecules and distort their electrophoretic mobilities.) (A/B good stored at RT for weeks; good for years at 4°c)

1.5 M Tris, pH 8.8

Suggest: 18.15 g Tris base Sigma T1503
50 ml glass distilled water; adjust to pH 8.8 with HCl
(with LPS, pH 8.3 works fine also)
q.s. 100 ml with glass-distilled water

0.5 M Tris, pH 6.8

Suggest: 3.0 g Tris base T1503
50 ml glass-distilled water, adjust to pH 6.8 with HCl
q.s. 100 ml with glass-distilled water
(The use of this in the stacking gels for LPS is not needed; instead use the 1.5 M Tris, above.)

Chamber Buffer, pH 8.3, 5X

Suggest Tris base (Sigma T1503) 52 gm
Glycine (Sigma G7403) 250 gm
SDS, Sodium dodecylsulfate (Sigma L4509) 26 gm
q.s. 1700 ml with glass-distilled water adjust to pH 8.3
q.s. 1736 ml with glass-distilled water

Dilute 5-fold with glass-distilled water for both top and bottom buffer chambers.

BioRad® Silver Stain Reagents

Silver Stain: cat #161-0445
Developer: cat #161-0447
(Don't use BioRad oxidizer! Use your own LPS oxidizer, per-iodic acid, next.)

LPS Oxidizer

220 ml glass-distilled water + 2.8 g per-iodic acid; dissolve.
+ 160 ml 95% ethanol + 20 ml acetic acid = 400 ml

An internal reaction occurs such that within a week this smell strongly of ethyl acetate (fingernail polish remover). Discard after one week. May be reused at least ten times.

C-M Tsai of NIH suggests the chemistry with LPS for this reagent. However, it is interesting to note that if this oxidizer is not used in the silver staining procedure, the results are nearly identical. This is supported by others who have shown that the periodate oxidation of the polysaccharide portion of LPS does not occur.