We use wash solution on the 2nd day of FISH experiment. It contains SSC + 3% Igepal , a non-denaturing detergent. Can you specify what is it's action on the slides?
Dear Smita,
the detergent applied to the cells on the slide dissolves membrane proteins and nuclear proteins making nucleic acids accessible for the FISH probes.
Best regards
Pavel
Dear Smita,
I agree with Mr Pavel. It make nucleic acids accessible for FISH probs. It dissolves links between proteins.
good luck
Fayza
Dear Smita,
In 1st day FISH ,Its function is same as said by Mr Pavel .
But in 2nd day FISH processing,we use Igepal /NP-40/tween-20 along with 2X SSC or 0.4XSSC to remove the unbound probe.The concentration of SSC and detergent detemines the stringency of washes.If concentration varies you might get false signals or non specific signals.
Can any one help on the stability of the solutions we use for FISH.. Ifwe are doing another set after two months, which all reagents can be used? Is prehyb buffer stable for two months? and also the after hybridization stringent wash solutions (varing conc. of SSC and NP40)
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