小鼠腫瘤壞死因子α(TNF-α)ELISA試劑盒子科生物研發生產,深圳子科生物ELISA試劑盒優質供應商,擁有10年的研發和生產經驗,擁有*的儀器設備,專業的技術人員和*的實驗技術,目前已經建立強大的實驗技術服務平臺,品牌設立為:ZIKER 。
小鼠腫瘤壞死因子α(TNF-α)ELISA試劑盒●品牌:子科生物ZIKER,美國R&D,美國immonoway,美國sciencell,德國IBL
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小鼠腫瘤壞死因子α(TNF-α)ELISA試劑盒標本的保存
一般說來,在 5 天內測定的標本可放置于 4℃,超過一周測定的需低溫凍存。 對收集后當天進行檢測的標本,儲存在 4 ℃ 備用,如有特殊原因需要周期收集標本, 將標本及時分裝后放在 -20 ℃ 或 -70 ℃ 條件下保存。避免反復凍融。 標本 2 -8 ℃ 可保存 48 小時, -20 ℃ 可保存 1 個月。 -70 度可保存 6 個月。部分標 本也可加入適當防腐劑,激素類標本需添加抑肽酶。
小鼠腫瘤壞死因子α(TNF-α)ELISA試劑盒標本的處理
可用作 ELISA 測定的標本十分廣泛,體液(如血清)、分泌物(唾液)和排泄物(如尿液)、 細胞培養上清、細胞、組織等均可作標本以測定其中某種成份。有些標本可直接進行測定,有些 則需經預處理。
● 血清:
室溫血液自然凝固 10-20 分鐘后,離心 20 分鐘左右( 2000-3000 轉 / 分)。收集上清。如 有沉淀形成,應再次離心。
● 血漿:
應根據試劑盒的要求選擇 EDTA 、檸檬酸鈉或肝素作為抗凝劑,加入 10 %( v/v )抗凝劑 ( 0.1M 檸檬酸鈉或 1% heparin 或 2.0%EDTA.Na2) 混合 10-20 分鐘后,離心 20 分鐘左右 ( 2000-3000 轉 / 分)。仔細收集上清。如有沉淀形 成,應再次離心。
●尿液、胸腹水、腦脊液:
用無菌管收集。離心 20 分鐘左右( 2000-3000 轉 / 分)。仔細收集上清。如有沉淀形成, 應再次離心。
●細胞培養上清:
檢測分泌性的成份時,用無菌管收集。離心 20 分鐘左右( 2000-3000 轉 / 分)。仔細收集 上清。檢測細胞內的 成份時,用 PBS ( PH7.2-7.4 )稀釋細胞懸液,細胞濃度達到 100 萬 /ml 左右。通過反復凍融,以使細胞破壞并放出細胞內成份。離心 20 分鐘左右( 2000-3000 轉 / 分)。仔細收集上清。保存過程中如有沉淀形成,應再次離心。
●細胞:
檢測細胞的成份時,對于貼壁細胞,去除培養液,用PBS、理鹽水或無血清培養液洗一遍。加 入適量裂解液。用槍吹打數下,使裂解液和細胞充分接觸。通常裂解液接觸細胞10 秒后,細胞 就會被裂解。對于懸浮細胞,離心收集細胞,用PBS、理鹽水或無血清培養液洗一遍.加入適 量裂解液, 用槍吹打把細胞吹散。用手指輕彈以充分裂解細胞。充分裂解后應沒有明顯的細胞沉 淀。如果細胞量較多,必需分裝然后再裂解。充分裂解后,10000-14000g離心3-5 分鐘,取上 清,即可進行后續操作。
●組織標本:
切割標本后,稱取重量。加入一定量的 PBS 或組織蛋白萃取試劑,緩沖液中可加入蛋白酶抑制 劑。用手工或勻漿器將標本勻漿充分。離心 20 分鐘左右( 2000-3000 轉 / 分)。仔細收集 上清置于 -20 度或 - 70 度保存,如有必要,可以將樣品濃縮干燥。分裝后一份待檢測,其余冷凍備用。
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