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脫脂奶(nai)粉（Westren Blot）是將鮮牛奶脫去脂肪再干燥而成，脂肪可降低1％左右，其他奶粉成份變化不大。

生產工藝：脫脂奶粉（Westren Blot）的生產工藝流程與全脂乳粉相類似，只是多了一個原料乳脫脂的過程。按熱處理程度進行分類，分類指(zhi)標為乳清蛋(dan)白氮(dan)指(zhi)數（WPNI），乳粉中(zhong)未變(bian)性乳清蛋(dan)白的量與(yu)熱處理程度成反比(bi)。

低熱(re)粉(fen)(fen)（WPNI>6mg/g乳(ru)粉(fen)(fen)）—72℃/15s或90℃/瞬時

中(zhong)熱粉(fen)（1.5mg/g）

三、western blot中脫脂(zhi)奶粉的作用

1，封閉

原理：用非(fei)抗原的蛋白(bai)去封閉膜的蛋白(bai)質結合位點(dian)，避免標記抗體固定在膜上引起背景增高，常用的封閉液有BSA,脫(tuo)(tuo)脂(zhi)奶(nai)粉,明膠, tween-20等,一般脫(tuo)(tuo)脂(zhi)奶(nai)粉。

實驗步(bu)驟(zou)：

5%脫(tuo)脂奶粉（用1x TBS配制(zhi)(zhi)，或1xTBST配制(zhi)(zhi)）封(feng)閉一晚上 或室溫1~2小時(shi)

2，一抗(kang)、二(er)抗(kang)孵育

原理(li)： 一抗(kang)(kang)可以(yi)與(yu)要檢測的(de)蛋白(bai)特異結合，二(er)抗(kang)(kang)可與(yu)一抗(kang)(kang)結合，同時二(er)抗(kang)(kang)上(shang)帶有特異的(de)酶(mei)（如(ru)辣根過(guo)氧化(hua)(hua)物酶(mei)HRP，堿(jian)性磷酸酶(mei)法AP等 ),標記(ji)酶(mei)可以(yi)分解(jie)化(hua)(hua)學(xue)顯(xian)色底物產生有顏色的(de)條帶或催(cui)化(hua)(hua)化(hua)(hua)學(xue)發光底物使膠片感光。

實驗步驟(zou)：

一抗(kang)(kang)孵育：5%脫脂(zhi)奶粉稀釋一抗(kang)(kang)到合適的濃度(du)(du)，與膜室溫孵育1小時或4度(du)(du)一晚上(shang)（抗(kang)(kang)體(ti)(ti)濃度(du)(du)及(ji)孵育時間需根據不同的抗(kang)(kang)體(ti)(ti)做相應調整， 1xTBST，洗(xi)膜3次，每次5分鐘。

二抗(kang)孵育：5%脫脂(zhi)奶(nai)粉稀釋二抗(kang)到合適的濃度，與膜(mo)室溫孵育1小時， 1x TBST,洗膜(mo)3次，每次5分鐘。

四、相關產品

C8070 Catalase   過氧化氫(qing)酶(mei)（牛(niu)肝(gan)）  1g

C8080 Cytochalasin B  細胞松(song)弛素B  1mg

 C8650 Cyclophosphamide monohydrate環(huan)磷(lin)酰(xian)胺  1g

C8100 DRISELASW 崩(beng)潰酶 ; 500mg

C8110 ConA Type A IV 刀(dao)豆蛋白A IV  25mg

C8120 Cytochromes C  細(xi)胞色素C(馬心)  25mg

C8130 Cytochromes C  細胞色(se)素C（牛心）    25mg

TEVDAB-1000U TEV蛋白酶 solarbio 1000u

C8140 CollagenaseⅠ  膠(jiao)原酶(mei)Ⅰ  100mg

C8150 CollagenaseⅡ  膠原酶Ⅱ  100mg

C8490 CollagenaseIII 膠原III 進口分裝 100mg

C8160 CollagenaseⅣ  膠(jiao)原(yuan)酶Ⅳ  100mg

C8170 CollagenaseⅤ  膠原酶Ⅴ  100mg

相關文獻：

《Effect of 1α, 25-dihydroxyvitamin D3 on the osteogenic differentiation of human periodontal ligament stem cells and the underlying regulatory mechanism》 作者:Yawen Ji, Panpan Zhang ,Yixiao Xing ,Linglu Jia ,Yunpeng Zhang, Tingting Jia ,Xuan Wu ,Bin Zhao, Xin Xu 期刊:International Journal of Molecular Medicine 影響(xiang)因子(zi):2.928 PMID:30365053

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