ELISA實(shí)驗(yàn)過程組織及細(xì)胞上清標(biāo)本的收集及處理

Tissue homogenates （組織標(biāo)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   （細(xì)胞培養(yǎng)上清液標(biāo)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.