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ISH - ethanol dehydration of whole mount embryos


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#1 miR cat

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Posted 17 May 2011 - 09:55 AM

Hi,
Can someone please tell me why NBT/BCIP signal bleaches out/vanishes after I dehydrate my embryos using ethanol?
How can I prevent this? Is dehydration even necessary?
Grateful for any advice on this.
Best,

#2 mdfenko

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Posted 19 May 2011 - 06:56 AM

the insoluble, colored, deposit formed when detecting with nbt/bcip is soluble in alcohol.
talent does what it can
genius does what it must
i do what i get paid to do

#3 miR cat

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Posted 19 May 2011 - 08:47 AM

Yes, I know that some component of the dye is solubilized and the colour changes from purple to blue, but ALL colour shouldn't vanish, especially since usually I watch the colour develop for over 5 hours after adding nct/bcip.




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