4.5. Measurement of Pro-angiogenic Factors in Tumor Samples from Patients and PDX Models
This protocol is extracted from research article:
Anti-Angiogenic Treatment in Pseudomyxoma Peritonei—Still a Strong Preclinical Rationale
Cancers (Basel), Jun 5, 2021; DOI: 10.3390/cancers13112819

Concentrations of pro-angiogenic factors in patient and PDX samples were quantified using two ELISA platforms. The electrochemiluminescence detection with Meso Scale Discovery (MSD) V-PLEX Angiogenesis Panel 1 Human Kit (VEGF-A/-C/-D, PlGF, bFGF/FGF2, sflt1/VEGFR1, Tie2) was performed according to the manufacturer’s protocol (MSD, Hercules, CA, USA). In addition, human pro-angiogenic factors VEGFA, PlGF and angiopoietin1 and mouse VEGFA and PlGF were analyzed with specific ELISA plates according to the manufacturer’s protocol (R&D Systems, Inc, Minneapolis, MN, USA). After thawing, samples were handled as described above and centrifuged briefly at 2000× g for 2 min at 4 °C. The supernatants were incubated on MSD V-plex or the specific ELISA plates. MSD V-plex plates were washed and read using SECTOR Imager 2400 software (MSD) and the ELISA (R&D Systems, Inc) on a plate reader Victor 1420 Multilabel Counter, (Wallac/PerkinElmer Life Sciences, Turku, Finland) at 450 nm with wavelength correction at 570 nm.

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