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solarbio RPMI1640 細(xì)胞培養(yǎng)試劑

紅細(xì)胞裂解液是一種比較溫和的紅細(xì)胞去除方法。既不損傷有核細(xì)胞又能充分的去除紅細(xì)胞。經(jīng)紅細(xì)胞裂解液裂解得到的組織細(xì)胞中不含紅細(xì)胞，可進(jìn)一步用于原代細(xì)胞培養(yǎng)、細(xì)胞融合、核酸與蛋白的分離和提取等。

紅細(xì)胞裂解的原理

紅細(xì)胞表面上有紅細(xì)胞專屬的表面抗原，當(dāng)裂解液中含可以攻擊特定紅細(xì)胞表面細(xì)胞抗原的酶的時候，就只會造成紅細(xì)胞的變形，生物通道擴(kuò)大、膨脹、裂解，而不會攻擊其他細(xì)胞。另外紅細(xì)胞有自己的電負(fù)性、滲透脆性、懸浮穩(wěn)定性、這都是區(qū)別于其他種類細(xì)胞的區(qū)別點。紅細(xì)胞裂解液就是利用這些特性裂解紅細(xì)胞的。

紅細(xì)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(xì)胞裂解液。如1ml新鮮全血加入3ml紅細(xì)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(xì)胞裂解后，溶液應(yīng)該是清亮透明的。

3. 收集細(xì)胞：4℃，450×g離心10分鐘以沉淀白細(xì)胞，小心吸棄上清液。

4. 向白細(xì)胞沉淀中加入兩倍體積的紅細(xì)胞裂解液，輕輕渦旋充分重懸白細(xì)胞。如起始血液為1ml，則加入2ml的紅細(xì)胞裂解液。

5. 4℃，450×g離心10分鐘沉淀白細(xì)胞，小心并*吸去上清液。

6. 重懸細(xì)胞，用于后續(xù)實驗；如提取RNA，于此步開始使用DEPC水配制的溶液進(jìn)行操作。

紅細(xì)胞裂解液的配置方法

取100 μl抗凝全血，加2 ml應(yīng)用液3#，混勻15～20 s；加2 ml應(yīng)用液2#，混勻15～20 s；加4 ml應(yīng)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(xì)胞懸液，備用。（這種紅細(xì)胞裂解法滿足了白細(xì)胞流式細(xì)胞儀檢測分選要求，價格低廉，是一種簡便快捷、經(jīng)濟(jì)實用的白細(xì)胞純化分選方法。）

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solarbio RPMI1640 細(xì)胞培養(yǎng)試劑