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problems with X-Gal stain


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#1 Mo_Mo

Mo_Mo

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Posted 19 May 2011 - 01:08 AM

Hi guys,

I need help with X-Gal stainings? The problem is that my staining is very weak, although I am incubating for quite some time. Does anyone have a suggestion?

This is what I do:

tissue preparation: unfixed murine skin embedded and snap-frozen in OCT, 18 µm sections stored at -20°C

for the staining: let section thaw, fix for 10 min with 4% formalin at RT, wash with PBS, add staining solution (conatins mgcl2, K3Fe(CN)6, K4Fe(CN)6, NP-40) including X-Gal (final conc. = 1 mg/ml), incubate O/N at 37°C

Thanks a lot.




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