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FFPE = formalin-fixed, paraffin-embedded; H&E = hematoxylin and eosin; IHC = immunohistochemistry; LCM = laser capture microdissection; OPN = osteopontin; PBS = phosphate-buffered saline; PCR = polymerase chain reaction; qPCR = real-time, quantitative PCR; RT = reverse transcriptase; TSP-1 = thrombospondin-1; TYRP-1 = tyrosinase-related protein-1.
Monoclonal antibodies to OPN, to TYRP-1 and to TSP-1 were purchased commercially (OPN [1:100], Chemicon International, Inc., Temecula, CA, USA; TYRP-1 [1:80, clone G-17], Santa Cruz Biotechnology, Inc., CA, USA; TSP-1 [1:320], BD Transduction Laboratories, CA, USA).
TSP-1 staining was positive in two of nine primary carcinomas without metastases (22%).
The efficacy and specificity of OPN, TSP-1 and TYRP-1 antibodies in IHC was tested by analysis of frozen tissue xenografts of known target gene expression [ 2 5 ] . Positive cytoplasmic staining was detected with the OPN antibody in nine of 26 (35%) primary breast carcinomas with nodal metastases.
The precise biological role of TSP-1 in human tumor progression awaits more definitive study.