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The Human VEGF-A Quantikine ELISA kit (R&D Systems) was used to measure the concentration of VEGF-A in plasma and tumor tissues from CRC patients according to the manufacturer’s instructions. To measure VEGF-A in tissue homogenates, we homogenized 150 mg of tumor tissues with Precellys Lysing Kit in 350 μl of phosphate-buffered saline with 1% protease inhibitor, followed by centrifugation for obtaining supernatants.
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