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now: Fri Sep 17 05:09:15 2021 ... mod: Sun Apr 4 18:42:43 2021
Zinc Negative Stain
This procedure is rapid and sensitive, able to detect 5 - 10 ng protein per band. The proteins are not themselves stained but show as clear transparent bands on an otherwise solid white gel background.
- After SdsPage rinse the gel in distilled water for 30 seconds.
- Incubate the gel in 0.2 M imidazole, 0.1% SDS for 15 minutes.
- Discard the previous solution and incubate the gel in 0.2 M zinc sulphate until the gel background becomes a deep white consistency, about 30 seconds
- protein bands will be transparent and colourless
- Stop the stain by rinsing with copious distilled water.
- be careful not to overstain the gel
- Take a picture of the gel by placing it on a dark background exposed to white light.
- Proteins are immobilized in the stain if the gel is kept hydrated. Stable for years.
- If dried, the stain will fade. Rehydration will restore the stain.
- Stained gels do not transfer well in semi-dry blotting apparatus. In submerged transfer apparatus, stained gels will transfer well if a zinc chelator like glycine is added eg: 25 mM Tris-HCl, 192 mM glycine pH 8.3 transfer buffer.
Castellanos-Serra L, Proenza W, Huerta V, Moritz RL, Simpson RJ. Electrophoresis 20:732-737 (1999)