I need to determine the drug content in dermal and subcutaneous region of skin...
I used it to separate epidermis from cutis in C57BL/6 mice (according to the protocol by prof. Ralf Paus)
1. You separate the back skin on the level of subcutis, with scissors, then cut the whole back skin and spread it on a sheet of white cardboard.
2. Punch round circles of skin (diameter ca 3-4 mm) with a skin punch for skin biopsies and a hammer.
3. put immediately to cold PBS with 1M NaBr and 1 mM PMSF
(phenylmethylsulfonyl fluoride),
4. Keep overnight in ice.
5. Then remove and separate epidermis from cutis using a rubber (such as the one on the blunt end of a pencil)
6. Keep sterile (wash 2x in 380% ethanol, and 2 times in sterile PBS)
Search for "salt split skin" in www.
This is from one of the publications:
...Salt split technique
Punch biopsy samples were incubated in 5 mL of NaCl (1 mol/L) at 4ºC for 24 h. The epidermis was then teased from the dermis with the use of a fine forceps....
If you plan to culture cells, use methods developed in the Yuspa laboratory. In this paper I give details
Cellular Populations Isolated from Newborn Mouse Skin Including Mesenchymal Stem Cells.
Lauren Kimlin, Victoria Virador
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