Evaluation of two sample preparation methods for prostate proteome analysis

For laboratory techniques that require well-preserved proteins, such as 2-DE, fresh tissue must be harvested and processed as fast as possible to avoid proteolytic degradation. We describe a modified method for harvesting tissue from radical prostatectomy specimens for proteome analysis and compare it with the standard technique. Cells were scraped from cut surfaces of 11 prostate specimens. A fraction of the material was smeared on a glass slide and Giemsa stained for morphological control. The sample was collected in a medium with protease inhibitors, and the protein material was prepared for 2-DE. Filtering and Percoll centrifugation were omitted. Sample locations were noted on a specimen map. From the same area, a tissue block was harvested for comparison. The block was processed with the conventional technique including mechanical disintegration, filtering and Percoll centrifugation. Quality measures of 2-DE were similar with both methods. With the scrape sampling technique, control smears showed abundant epithelial cells and a cleaner background and processing was faster than with tissue block sampling. For proteomic analysis, the scrape sample technique has several advantages over the tissue block method.

Hellman, UlfUppsala universitet,Ludwiginstitutet för cancerforskning(Swepub:uu)ulfhm (författare)
Palmberg, CarinaKarolinska Institutet (författare)
Auer, GertKarolinska Institutet (författare)
Hellström, MagnusKarolinska Institutet (författare)
Franzén, BoKarolinska Institutet (författare)
Jörnvall, HansKarolinska Institutet (författare)
Egevad, LarsKarolinska Institutet (författare)
Uppsala universitetLudwiginstitutet för cancerforskning (creator_code:org_t)

Av författaren/redakt...: Lexander, Helena; Hellman, Ulf; Palmberg, Carina; Auer, Gert; Hellström, Magnu ...; Franzén, Bo; visa fler...; Jörnvall, Hans; Egevad, Lars; visa färre...

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