Cd51 fitc

Blood samples were obtained after 12 hours of fasting and the analyses were
performed at the central laboratory of our university. All athletes were
allowed to maintain their daily exercises program even on the day before
blood sample collection. The athletes had very similar exercise training
programs, corresponding to two long-distance running sessions every day, 15
km in the morning and 10 km in the afternoon, and intensive training
(100-1,000 meter shots, repeated many times)twice a week, on Tuesday and
Thursday mornings. All blood samples were collected on Thursdays, before
exercise.
Measurements of EPCs and MPs were performed as previously reported, using
fresh blood samples in EDTA containing tubes.^{12 (link)-15 (link)}For determination of EPCs, a minimum of 500,000 events was acquired by
flow-cytometry (FACSCalibur, BD Biosciences, USA). Fluorescently labeled
mouse anti-human antibodies were used for EPCs (CD34 FITC, BD Biosciences,
USA; CD133 APC, Miltenyi Biotec, USA; KDR PE, R&D Systems, USA), PMPs
(CD42 FITC and CD31 PE, BD Biosciences, USA) and EMPs (CD51 FITC, BD
Biosciences). Disposable containers (BD Biosciences) were used to quantify
the number of microparticles per microliter of
platelet-poor plasma (PPP).

Circulating endothelium-, monocyte- and platelet-derived microparticles were measured as previously reported (21 (link),22 (link)). In brief, blood samples were collected and centrifuged (160g; 20-22°C; 10 min) to obtain platelet-rich plasma (PRP). The PRP was centrifuged (1500g; 20-22°C; 6 min) to obtain platelet-poor plasma (PPP). Furthermore, PPP (50 µL) was labeled (20 min; room temperature) with CD51FITC, CD42FITC and CD31 PE, and CD14FITC (BD Biosciences) for the identification of endothelial, platelet, and monocytic microparticles, respectively. Isotypes (BD Biosciences) were used as controls. Microparticles were quantified per microliter of PPP injected into the cytometer, according to the standard protocol. TruCOUNT (BD Biosciences) tubes containing a known number of beads were used to quantify the number of microparticles per microliter of PPP.