产品介绍：
阿拉丁生产的脱脂奶粉(Blotting Grade)，即nonfat powdered milk, non-fat powdered milk, nonfat dry milk or skim milk for western blotting applications，是通过将精选的低免疫源性的鲜牛奶脱去脂肪后干燥而成，并经Western检测验证可以用于配制Western检测时的封闭液(Blocking Buffer)、一抗稀释液和二抗稀释液。脱脂奶粉是最常用的Western检测的封闭液组分之一。本产品溶解性好，使用本产品封闭后，可以有效减少一抗或二抗和膜的非特异性结合，降低背景，增强信噪比。脱脂奶粉配制的封闭液和BSA(牛血清白蛋白)配制的封闭液不同，前者是不透明的，后者是透明的。更多的用户喜欢使用BSA作为主要的有效封闭成分。但BSA的价格远高于脱脂奶粉。对于特定目标蛋白的Western检测，有时使用BSA作为主要封闭组分的效果更好，有时使用脱脂奶粉的封闭效果更好。通常优先参考一抗生产商推荐的封闭液进行封闭，如果封闭效果不好，可以尝试BSA和脱脂奶粉中的另外一种。如果希望快速有效地完成封闭和检测，推荐使用阿拉丁生产的UltraBio™系列产品，包括UltraBio™Western封闭液、UltraBio™ Western一抗稀释液和UltraBio™ Western二抗稀释液等多个产品。UltraBio™系列产品能实现超快速并且更好效果的的封闭。
注意事项：
通常在室温封闭60分钟即可。如果对于一些背景非常高的抗体，可以尝试4℃封闭过夜。脱脂奶粉成分比较复杂，可能含有微量的生物素，故不适合生物素-亲和素检测系统。尽量避免用于稀释anti-goat及anti-sheep的二抗。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。
使用说明：
1.配制Western封闭液：脱脂奶粉的浓度通常为3-5%(w/v)，推荐使用阿拉丁生产的 E616323 Western洗涤液或者稀释成1×的进行配制，也可以使用TBSTw (20 mM Tris pH 7.6, 137 mM NaCl, 0.1% Tween 20)进行配制。
2.一抗和二抗也可以用该封闭液稀释。
3.样品完成转膜后，用Western洗涤液洗涤蛋白膜1-2分钟。加入Western封闭液，封闭60分钟后，然用Western洗涤液洗涤蛋白膜1-2次，每次1-2分钟。随后进行一抗孵育等后续操作。
详细的Western操作可以参考如下的相关网页： https://www.aladdin-e.com/zh_cn/faqs/western-blot-protocol.html
Aladdin Blotting Grade Nonfat Dry Milk, also known as nonfat powdered milk, non-fat powdered milk or skim milk, is specially formulated for Western blotting. It is produced by defatting carefully selected fresh milk with low immunogenicity followed by drying. Validated for Western blot assays, this product can be used to prepare blocking buffer, primary antibody diluent and secondary antibody diluent. Nonfat dry milk is one of the most commonly used components for Western blot blocking solutions. This product features excellent solubility. When used for blocking, it effectively reduces non-specific binding between antibodies and blotting membranes, lowers background signals and improves signal-to-noise ratio. Blocking buffer prepared with nonfat dry milk appears opaque, while that formulated with BSA (Bovine Serum Albumin) is clear. Many researchers prefer BSA as the main blocking agent, yet BSA is considerably more expensive than nonfat dry milk. For Western blot detection of specific target proteins, either BSA or nonfat dry milk may yield superior blocking results depending on the experimental conditions. It is recommended to follow the blocking buffer guidelines provided by the primary antibody manufacturer in the first place. If unsatisfactory blocking performance occurs, switch to the alternative blocking agent. For rapid and efficient blocking and detection, Aladdin’s UltraBio™ product series is highly recommended, including UltraBio™ Western Blocking Buffer, UltraBio™ Primary Antibody Diluent, UltraBio™ Secondary Antibody Diluent and other related products. The UltraBio™ series enables ultra-fast blocking with optimized performance.
Precautions:
Standard blocking is performed at room temperature for 60 minutes. For antibodies that cause extremely high background, overnight blocking at 4 °C is recommended. Nonfat dry milk has a complex composition and may contain trace biotin, so it is not suitable for biotin-avidin detection systems. Avoid using it to dilute anti-goat and anti-sheep secondary antibodies. This product is intended for scientific research use only by professionals. It shall not be used for clinical diagnosis, treatment, food or pharmaceutical production, and shall not be stored in residential premises. For personal safety and health, please wear a lab coat and disposable gloves during operation.
Instructions for Use:
1. Preparation of Western Blocking Buffer:
The concentration of nonfat dry milk is generally 3–5% (w/v). It is recommended to prepare with Aladdin Cat. No. E616323 Western Washing Buffer (diluted to 1× working solution) or with TBSTw (20 mM Tris, pH 7.6, 137 mM NaCl, 0.1% Tween 20).
2. This blocking buffer can also be used to dilute primary and secondary antibodies.
3. After membrane transfer:
Rinse the protein membrane with Western Washing Buffer for 1–2 minutes. Add Western Blocking Buffer and block for 60 minutes. Wash the membrane 1–2 times with Western Washing Buffer, 1–2 minutes each time. Proceed to subsequent steps such as primary antibody incubation.
For detailed Western blot procedures, please refer to the webpage below: https://www.aladdin-e.com/en_us/faqs/western-blot-protocol.html
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