I have never success a staining pax7, myoD, eMHY on mouse tissue. (DSHB antibody)
When I did a heat-retrieval, staining is success but background was too strong.
So I'm not sure if this is a good staining.
My protocol is next.
1. mouse limb was harvest and fix on 4% PFA overnight.
2. Sucrose 10%, 20%, 30%
3. OCT embedding
4. tissue section
5. Staining
a. OCT wash 3 times (1X PBS)
b. M.O.M kit 1drop/1ml 1h
c. 1%BSA + 0.3% Triton blocking and perm
d. 1st AB 4'C overnight.
e. wash 3 times and 2nd AB 2h RT.
f. Wash and DAPI. Mounting.
Please let me know the way I can solve this..
Thanks.
You may give the free-floating staining a try. You basically do all the staining steps where the sections are free in the solution and at the last stage, mount them on the slides.
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