海角国精产品三区二区三区,91精品国产综合久久福利,国产精品无码亚一区二区三区,国产成人精品免费视频www,偷拍视频对白不卡高清精品

10×酸性蛋白Native PAGE轉膜緩沖液

簡介：

10×酸性蛋白Native PAGE轉膜緩沖液適用于酸性蛋白電泳后凝膠上非變性酸性蛋白的轉膜。本轉膜液配制便捷，稀釋后無需調節pH值，pH約為9.2。緩沖液主要成分為Tris，甘氨酸和微量SDS。該緩沖液最終可配成10升即用型1×緩沖液(ye)（要補(bu)加甲醇）。

保存條件：

粉末(mo)裝常溫保存(cun)，兩(liang)年有效。

配制方法：

將10×轉膜緩沖液粉末全部溶解于1升超純水中，徹底溶解，即配成10×轉膜緩沖液。根據下表配制成1×即(ji)用型轉膜緩沖液(ye)

使用方法：

1. 酸性蛋白非變性電泳：

各種蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)質分子由(you)于(yu)所含的(de)(de)(de)堿(jian)(jian)性(xing)(xing)(xing)氨(an)(an)基酸(suan)(suan)(suan)(suan)和(he)酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)氨(an)(an)基酸(suan)(suan)(suan)(suan)的(de)(de)(de)數目不同，因而有各自(zi)(zi)的(de)(de)(de)等(deng)電(dian)(dian)(dian)(dian)(dian)點。凡堿(jian)(jian)性(xing)(xing)(xing)氨(an)(an)基酸(suan)(suan)(suan)(suan)含量(liang)較多(duo)(duo)的(de)(de)(de)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)質稱為堿(jian)(jian)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)，等(deng)電(dian)(dian)(dian)(dian)(dian)點偏堿(jian)(jian)性(xing)(xing)(xing)，pI＞7，如組蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)、精蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)等(deng)。反之，凡酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)氨(an)(an)基酸(suan)(suan)(suan)(suan)含量(liang)較多(duo)(duo)的(de)(de)(de)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)質，等(deng)電(dian)(dian)(dian)(dian)(dian)點就偏酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)，pI＜7，自(zi)(zi)然界中(zhong)大多(duo)(duo)數蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)都(dou)是酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)。分離堿(jian)(jian)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)時(shi)(shi)候，要(yao)利用低 pH 凝(ning)膠(jiao)(jiao)系(xi)統，分離酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)時(shi)(shi)候，要(yao)利用高 pH 凝(ning)膠(jiao)(jiao)系(xi)統。酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)通常在非變性(xing)(xing)(xing)凝(ning)膠(jiao)(jiao)電(dian)(dian)(dian)(dian)(dian)泳(yong)(yong)中(zhong)采用的(de)(de)(de) pH 是 8.8的(de)(de)(de)緩沖系(xi)統，蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)會(hui)帶(dai)負電(dian)(dian)(dian)(dian)(dian)荷，蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)會(hui)相陽(yang)極移(yi)動；而堿(jian)(jian)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)通常電(dian)(dian)(dian)(dian)(dian)泳(yong)(yong)是在微酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)環境(jing)下進行，蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)帶(dai)正(zheng)電(dian)(dian)(dian)(dian)(dian)荷，這時(shi)(shi)候需要(yao)將陰(yin)極和(he)陽(yang)極倒置才可以電(dian)(dian)(dian)(dian)(dian)泳(yong)(yong)。酸(suan)(suan)(suan)(suan)性(xing)(xing)(xing)蛋(dan)(dan)(dan)(dan)(dan)白(bai)(bai)電(dian)(dian)(dian)(dian)(dian)泳(yong)(yong)可參考使(shi)用非變性(xing)(xing)(xing)PAGE凝(ning)膠(jiao)(jiao)制備及電(dian)(dian)(dian)(dian)(dian)泳(yong)(yong)試劑盒（貨號:RTD6130）。

2. 轉膜：

 ① 轉膜方法：酸性蛋白在非變性狀態的轉膜建議用濕轉法。

 ② 三明治結構：由于酸性蛋白pI＜7 在堿性轉膜緩沖液中帶負電荷，轉膜三明治結構與傳統轉膜相同，即根據“黑膠白膜”制作三明治，即膜置(zhi)(zhi)于轉(zhuan)膜夾芯正(zheng)極一側(ce)，凝膠置(zhi)(zhi)于轉(zhuan)膜夾芯負極一側(ce)，這樣(yang)凝膠上(shang)帶負電荷的酸性蛋白才能轉(zhuan)移到(dao)膜上(shang)。

酸性蛋白轉膜三明治制(zhi)作：

負極（電轉夾黑色面）-海綿墊-3至5層濾紙-凝膠-膜-3至5層濾紙-海綿墊-正極（電轉夾白色面）

 ③ 膜的選(xuan)擇(ze)：膜可以用PVDF膜或NC膜，根據蛋白(bai)大小選(xuan)擇(ze)膜的孔(kong)(kong)徑。一般說來，大于(yu)20kD蛋白(bai)選(xuan)擇(ze)0.45μm孔(kong)(kong)徑，低于(yu)20kD選(xuan)擇(ze)0.22μm孔(kong)(kong)徑。PVDF膜使用前要(yao)用無水(shui)甲醇潤(run)濕(shi)活化，NC膜只需用轉膜緩沖液潤(run)濕(shi)就可以。

④ 轉膜條件：

由于在(zai)非變性(xing)條(tiao)件下(xia)，不同酸(suan)性(xing)蛋(dan)白的(de)空間結(jie)構，聚合狀態，電荷數(shu)量都(dou)有不同，以下(xia)轉(zhuan)膜條(tiao)件僅供參考，客(ke)戶針對自己的(de)目的(de)蛋(dan)白，最(zui)好經過1-2次預實(shi)驗后，確定最(zui)佳(jia)的(de)轉(zhuan)膜條(tiao)件。