[Histonet] Proteinase K treatment

Segovia, Claudia csegovia <@t> nsalabs.com
Thu Oct 3 16:40:17 CDT 2013


Hello histoneters!

This is my first time using the net. I hope to find here some answers to my problem. I am working with free floating sections embedded in gelatin. I need to use Proteinase K to detect alpha- synuclein aggregates in brain tissue. I tried concentrations in the range of 1 Ug/ml to 5Ug/ml, incubation time is 10 minutes at room temperature. I noticed that after several rinses the Proteinase K keeps being active, because the gelatin in the sections gets sticky  and even disappears overtime. I did several rinses (4 X 10 min each) and I am afraid that the antibody will get degraded in the overnight incubation. How can I stop this enzymatic reaction? 
I will truly appreciate any insights,
Thank you!

Claudia N. Segovia
Senior Neurohistologist
Antibody Specialist
NeuroScience Associates
10915 Lake Ridge Drive
Knoxville, TN  37934
865-675-2245
csegovia <@t> nsalabs.com

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