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.
Lexander
Helena
H
Department of Oncology and Pathology, Karolinska Hospital, Karolinska Institute, SE-171 76 Stockholm, Sweden.
Hellman
Ulf
U
Palmberg
Carina
C
Auer
Gert
G
Hellström
Magnus
M
Franzén
Bo
B
Jörnvall
Hans
H
Egevad
Lars
L
D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008297:Male; D009363:Neoplasm Proteins / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D020543:Proteome; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009363:Neoplasm Proteins; D020543:Proteome