Post subject: Background with FISH: pre-adsorption?
I'm using anti-DIG-POD with the TSA system on mouse skin sections, but having background (small spot-like) even if I hybridize without a probe. I'm using 2% blocking reagent + 10% donkey serum for blocking, incubate the anti-DIG at 1/100 for 1hr, wash 3x10mins with PBST and make the TSA reaction for 7mins and wash again.
Has anyone ever used something like a pre-adsorption for the Ab? Ie. incubating the Ab with unhybridized tissue, filtrate and use the solution for detection of DIG?
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