The procedures of c-Kit protein immunohistochemical staining were as follows: dewax
and dewater the sections; soak with PBS for 5 min; sodium citrate for restoration; natural cooling for 5 min; wash with cold water for 10 min; soak with PBS for 5 min; incubate with 3% hydrogen peroxide solution for 15 min; wash with PBS for 2 min; add rabbit anti-mouse c-Kit polyclonal antibody (Thermo Scientific, USA) and incubate for 1.5 h; wash with PBS for 2 min; add enzyme labeled second antibody for 30 min; wash with PBS for 2 min; add diaminobenzidine (DAB) colored solution for 10 min; wash with distilled water; hematoxylin for counterstaining for 5 min; dehydration of gradient ethanol for 5 min; dehydration of xylene for 5 min; wash with PBS for 3 times; and seal with neutral gum.
Sections were baked at 60[degrees]C for 60 min in a dehydration oven, then dewaxed
in Bond Dewax
solution, and rehydrated in Bond Wash solution (Leica Microsystems).
paraffin slices with routine dimethylbenzene and rinse them with gradient alcohol to hydration.
Routine immunolocalization procedures were used to dewax
and rehydrate the slides.
Using the automated protocol of the Leica Bond Rx Automated Stainer (Leica Products/Equipment, Leica Microsystems, Inc., Buffalo Groove, IL), the slides were baked for 30 minutes and dewaxed
with Leica Bond Dewax
solution (Cat #AR9222).
To remove paraffin from tissue sections before rehydration and immunostaining on the Bond automated system (Leica Biosystems Newcastle Ltd., UK), Bond Dewax
solution was used as previously reported .
The liver sections were dewaxed
in a Bond Dewax
solution and rehydrated in alcohol and a Bond Wash solution (Leica Microsystems Inc., USA).
Four-micrometer paraffin sections were dewaxed
in a Bond Dewax
solution, rehydrated in alcohol and Bond Wash solution (Leica Microsystems).
For high-quality nuclei isolation, we used the Insitus Biotechnologies kit, which was designed for paraffin-embedded tissue slides and bifunctional skip dewax
. Nuclei were resuspended in 200 [micro]L of fixative (3 parts methanol and 1 part glacial acetic acid).
At that point in time the opportunity to enter the financially attractive lube base oil market was born because the heaviest product from the hydrocracking unit is excellent feedstock to dewax
and manufacture very high quality lube base oils.
This is a time when biological and mechanical means are used to incorporate moisture, soften the straw, increase density (cause some shrink) and dewax