Pierce Clear Milk Blocking Buffer (10X) / 透明牛奶阻隔缓冲液（10X）
货号：37587
规格：100 mL
价格：1800
产品类型：免疫印迹/组化
品牌：Thermo Fisher
| Pierce 透明牛奶封闭缓冲液 (10X) 是 Western 印迹应用中自制脱脂乳封闭缓冲液的长有效期、高效替代物。我们的透明牛奶封闭缓冲液 (10X) 是一种预配制的牛奶溶液，澄清且稳定，与硝酸纤维素和 PVDF 膜配合使用时可用于封闭过量的非特异性结合位点、减少 Western 印迹中的背景信号以及稀释抗体。与基于溶解的无脂干奶的自制缓冲液相比，该缓冲液可提供更低的背景、更高的灵敏度、更长的有效期和可重现的结果。因为其含有一些内源性生物素，所以不建议用于基于亲和素的技术。 |
| 数据： |
Milk blocking buffer compatibility with nitrocellulose and PVDF membranesSerial dilutions of A549 cell lysate (5, 2.5, 1.25, 0.62, 0.31, 0.16 and 0.78μg) were prepared and separated by electrophoresis. The proteins were transferred to nitrocellulose (Part No. 88014) and PVDF membranes (Part No. 88585) and the membranes were blocked for 30 minutes with 5% skim milk in PBS + 0.05% Tween™20 or Clear Milk Blocking Buffer (Part No. 37587) prepared at 1X in ultrapure water. After blocking, the membranes were incubated with primary antibodies to targets A, B and C. The membranes were washed and then incubated with 10ng/mL of HRP-Conjugated Goat Anti-Mouse IgG (Part No. 31430) and 6.7ng/mL of HRP-Conjugated Goat Anti-Rabbit IgG (Part No. 31460) then washed again. Working solution of SuperSignal West Dura Extended Duration Substrate (Part No. 34076) was prepared according to the instructions and added to the membranes for 5 minutes. The membranes were removed from the substrate, placed in plastic sheet protectors and exposed to CL-XPosure Film (Part No. 34090) for 5 seconds.Antibodies:A. Heat Shock Protein 86 (Hsp86) (Part No. PA3-013) at 1μg/mLB. NFκB (Millipore No. 06-418; 1mg/mL) at 1μg/mLC. PP2A (Millipore No. 05-421) at 0.2μg/mL |
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