ELISA实验过程组织及细胞上清标本的收集及处理

Tissue homogenates （组织标本） - The preparation of tissue homogenates will vary depending upon tissue type. For this assay, tissues were rinsed in ice-cold PBS（0.02mol/L,pH 7.0-7.2） to remove excess blood thoroughly and weighed before homogenization. Minced the tissues to small pieces and homogenized them in 5-10 mL of PBS with a glass homogenizer on ice（Micro Tissue Grinders woks, too）. The resulting suspension was sonicated with an ultrasonic cell disrupter or subjected to two freeze-thaw cycles to further break the cell membranes. After that, the homogenates were centrifugated for 5 minutes at 5000×g. Remove the supernate and assay immediay or aliquot and store at ≤-20oC.

Cell culture supernates   （细胞培养上清液标本）  - Centrifuge samples for 20 minutes at 1000×g. Remove particulates and assay immediay or store samples in aliquot at -20oC or -80oC for later use. Avoid repeated freeze/thaw cycles.