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Home-made Anti-bleach (?), vectashield and Immunocytochemistry

ICC 2ndAb Vectashield

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#1 mordiano

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Posted 08 November 2012 - 04:30 AM

Hi all,

I have some ideas regarding immunocytochemistry.

They are more like questions actually.

Is there a good way to protect samples from bleaching? Samples with a bound flourescent secondary antibody?

I used to use vectashield in another lab but here everyone is using glycerol/PBS. Will there be a diffrence?

Can I add something to make it "vectashield-ish"?

From what I understtod vectashield are cheap chemicals sold expensively...



Cheers!

PS. And do I need to bother at all?

#2 bob1

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Posted 08 November 2012 - 01:35 PM

Keep the slides in the dark at 4 deg C or lower (frozen even). Slowfade and slowfade gold work well for me.

#3 mordiano

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Posted 12 November 2012 - 08:08 AM

Good idea, realised (checking, after hearing you) that 70% glycerol would not freeze at -4*C. Still though Im not sure about the integrity of my small worms :/

At any rate I´ll start looking at the company products you mentioned.

#4 El Crazy Xabi

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Posted 20 November 2012 - 10:31 PM

Sigma also have its own product: Fluoroshield
http://www.sigmaaldr...ng=en&region=AU

But as all of them are quite expensive. At least this one (I use it) is a mounting medium rather than just a antifading reagent. It may contain some polymer based on the plastic/silicone-like residue it left on the rim of the bottle.

I don't know about your worms but most fluorescent-stained stuff is recommended to be stored at -20° C rather than 4° C, though the later is common for short term storage: examination the next day or so.





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