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B-gal staining troubleshooting - No blue cells on positive controls

B-gal staining;

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#1 Irina_Vavi



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Posted 15 August 2014 - 02:57 AM

Hello everyone!

I have been doing B-gal staining on frozen sections without success. I use the SIGMA kit for senescent cells detection, which should be pretty straight forward, but I get no staining on my positive controls.

The protocol I follow is this one:

-Incubate x-gal sol at 37*C for one hour

-Defrost sections for 3 min

-Fix with 1X fixative sol (from kit) for 7 min

-Wash 3 times with 1x pBS

-Add staining mix and incubate overnight, at 37*C

-Countersatin with nuclear red


I don't know if a might be missing something or if my B-gal activity is being affected by a step in the protocol.




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