B. Krušlin

J. Matić

M. Ulamec

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242 Acta Clin Croat,  Vol. 49,  No. 2,  2010Acta Clin Croat 2010; 49:211-243 Conference PapersTHE IMMUNOHISTOCHEMICAL PROCEDUREFOR THE DETECTION OF MOUSE MONOCLONAL C-MYC EXPRESSIONJ. Matić, M. Ulamec, B. KrušlinLjudevit Jurak University Department of Pathology, Sestre milosrdnice University Hospital, Zagreb, Croatiac-Myc protein is a transcription factor encoded by the c-Myc gene located on human chromosome 8q24. c-Myc plays an important role in many physiological processes. Chromosomal translocations or amplifi -cations can induce c-Myc gene alterations. Several studies have shown that c-Myc gene alterations play an essential role in the development of tumor neovas-cularization. Th erefore, c-Myc is an object of numer-ous studies dealing with novel therapeutic approaches and/or prognostic factors in cancer treatment. Th e antibody to c-Myc is currently in use only for scien-tifi c purposes and there are no standardized recom-mendations for the detection of c-Myc. We present our experiences in this fi eld. Th e study was done on one hundred and fi fty paraffi  n blocks with prostate carcinoma from the Ljudevit Jurak Department of Pathology archive. Specimens were fi xed in 10% buff -ered formaldehyde, embedded in paraffi  n and cut at 3 μm for immunostaining in Dako Autostainer. PAP method was used with LSAB/HRP universal-Dako visualization and en vision FLEX.  Pretreatment of samples was done within PT link. Optimal dilution was 1:150. c-Myc antibodies from Abcam (9E10) and Santa Cruz (9E10) were used. We employed several diff erent techniques in the preparation of c-Myc im-munostained sections, according to protocols from various manufacturers. At the end we chose a protocol that was sensitive enough for the detection of c-Myc and its evaluation. c-Myc antibody is most frequently used for scientifi c purposes and there is a need for standardization of c-Myc detection procedure in rou-tine practice. Experiences from diff erent laboratories are valuable because they allow comparison and revi-sion of the procedure.

The Immunohistochemical Procedure for the Detection of Mouse Monoclonal C-Myc Expression

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