How to stain MW in nitrocellulose membran? - Western bloting (Nov/13/2007 )

What dye do you use to stain the protein MW transferred to the nitrocellulose membrane when Western bloting??

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medium sensitivity: Amido black
high sensitivity: India ink

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Indian ink????

are you talking about normal indian ink used for fountain pens????
How do you use it?

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reversible stain: ponceau s.

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yes, the trade name is "India ink" (Higgins black India ink, Eberhard Faber, Pelikan) dilute a drop of about 50 µl in 50 ml TTBS; maximum stain is reached after several hours, but after ~0.5h you may see a stain; persumably, colloidal carbon binds to proteins

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I agree to use ponceau to check to see:

i)Transfer of proteins from gel to Nitrocellulose membrane (transient as described, disappears with washing)
ii) Gives an idea of Equal sample loading.

However the MW weight markers we use are COLOURED so are easily observed after transfer and BEFOR ponceau staining.

Hope this is useful

Rhombus

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Thanks for your comments!

You are right, I wanted to buy prestained MW, unfortunately, the guy who worked here acquired lot of no-stained MW !! so, We have enough for al least two years…

The beared:
Thanks for the info,
I will try indian ink... after incubation with it, just wash with buffer, right?

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yes; if staining is not strong enough you can continue with incubation

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