{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,20]],"date-time":"2026-06-20T00:53:22Z","timestamp":1781916802606,"version":"3.54.5"},"reference-count":26,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2004,1,21]],"date-time":"2004-01-21T00:00:00Z","timestamp":1074643200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["pathsocjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Pathology"],"published-print":{"date-parts":[[2004,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    In recent years, laser capture microdissection (LCM) has been used successfully to obtain distinct populations of cells for subsequent molecular analysis. Because of the limited sample availability and the absence of\n                    <jats:italic>in vitro<\/jats:italic>\n                    amplification steps for proteins, the use of LCM for proteome analysis largely depends on highly sensitive protein detection methods. In this study, a western blot protocol was developed and validated for the detection of \u03b2\u2010actin and the moderately expressed cell death protein caspase\u20103 in small numbers of cells. Initially, cultured human U937 monocytes and whole sections of paraffin wax\u2010embedded, alcohol\u2010fixed human tonsils were used to optimize protein electrophoresis and western blotting conditions. High\u2010performance NuPAGE Bis\u2010Tris gels in combination with high\u2010quality transfer membranes, optimized antibody concentrations, and a sensitive chemiluminescent substrate provided a strong signal for \u03b2\u2010actin with \u223c500 U937 cells. In the same way, procaspase\u20103 could be identified with \u223c1000 cells. Similar results were obtained with germinal centre cells that were procured from paraffin wax\u2010embedded, alcohol\u2010fixed human tonsils by LCM. Treatment of U937 cells with etoposide rapidly induced cell death and allowed the detection of active caspase\u20103 with \u223c2500 cells (0.8 pg of protein). The findings of this study suggest that western blotting is a valuable adjunct to proteome analysis of LCM procured cells. Copyright \u00a9 2004 Pathological Society of Great Britain and Ireland. Published by John Wiley &amp; Sons, Ltd.\n                  <\/jats:p>","DOI":"10.1002\/path.1525","type":"journal-article","created":{"date-parts":[[2004,2,26]],"date-time":"2004-02-26T15:24:06Z","timestamp":1077809046000},"page":"382-388","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Western blot analysis of a limited number of cells: a valuable adjunct to proteome analysis of paraffin wax\u2010embedded, alcohol\u2010fixed tissue after laser capture microdissection"],"prefix":"10.1002","volume":"202","author":[{"given":"Wim","family":"Martinet","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanessa","family":"Abbeloos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathalie","family":"Van Acker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guido 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