目錄:北京索萊寶科技有限公司>>細胞培養相關>>細胞培養試劑>> 10491solarbio RPMI1640 細胞培養試劑
供貨周期 | 現貨 | 規格 | 500ml |
---|---|---|---|
貨號 | 10491 | 應用領域 | 醫療衛生,化工,生物產業,石油 |
主要用途 | 細胞培養試劑 |
solarbio RPMI1640 細胞培養試劑
紅細胞裂解液是一種比較溫和的紅細胞去除方法。既不損傷有核細胞又能充分的去除紅細胞。經紅細胞裂解液裂解得到的組織細胞中不含紅細胞,可進一步用于原代細胞培養、細胞融合、核酸與蛋白的分離和提取等。
紅細胞裂解的原理
紅細胞表面上有紅細胞專屬的表面抗原,當裂解液中含可以攻擊特定紅細胞表面細胞抗原的酶的時候,就只會造成紅細胞的變形,生物通道擴大、膨脹、裂解,而不會攻擊其他細胞。另外紅細胞有自己的電負性、滲透脆性、懸浮穩定性、這都是區別于其他種類細胞的區別點。紅細胞裂解液就是利用這些特性裂解紅細胞的。
紅細胞裂解液的使用說明
1. 1倍體積的新鮮全血,加入3倍體積的紅細胞裂解液。如1ml新鮮全血加入3ml紅細胞裂解液,輕輕渦旋或顛倒混勻。
2. 冰上放置15分鐘,其間輕輕渦旋混勻兩次,紅細胞裂解后,溶液應該是清亮透明的。
3. 收集細胞:4℃,450×g離心10分鐘以沉淀白細胞,小心吸棄上清液。
4. 向白細胞沉淀中加入兩倍體積的紅細胞裂解液,輕輕渦旋充分重懸白細胞。如起始血液為1ml,則加入2ml的紅細胞裂解液。
5. 4℃,450×g離心10分鐘沉淀白細胞,小心并*吸去上清液。
6. 重懸細胞,用于后續實驗;如提取RNA,于此步開始使用DEPC水配制的溶液進行操作。
紅細胞裂解液的配置方法
取100 μl抗凝全血,加2 ml應用液3#,混勻15~20 s;加2 ml應用液2#,混勻15~20 s;加4 ml應用液1#,混勻15~20 s;300×g離心2 min,棄上清;收集沉淀,加PBS緩沖液,制成白細胞懸液,備用。(這種紅細胞裂解法滿足了白細胞流式細胞儀檢測分選要求,價格低廉,是一種簡便快捷、經濟實用的白細胞純化分選方法。)
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solarbio RPMI1640 細胞培養試劑