Western溶液套装(封闭液、洗涤液、一抗和二抗稀释液)

阿拉丁生产的Western溶液套装(封闭液、洗涤液、一抗和二抗稀释液)，是一种包含了Western Blot实验过程中常用的封闭液、洗涤液、一抗和二抗稀释液的溶液套装。本产品中的一抗稀释液(Primary Antibody Dilution Buffer for Western Blot) 可以用于Western时一抗(primary antibody)的稀释和配制。其主要有效成分为经过反复优化的适量BSA、适当的去垢剂和一抗稳定试剂等，一方面能有效减少一抗的非特异性结合，另一方面能有效提升稀释后一抗的稳定保存时间。使用本Western一抗稀释液稀释和配制的一抗可以在4℃保存和使用不少于六个月，可以反复多次用于Western，节约您宝贵的抗体。本产品中的Western封闭液(Blocking Buffer for Western Blot) 可以用于Western时转移了蛋白样品的PVDF膜或NC膜的封闭。其主要有效成分为经过反复优化的适量BSA和适当的去垢剂等，一方面能有效封闭膜上的蛋白结合位点，另一方面能有效减少后续一抗和二抗的非特异性结合。使用本Western封闭液封闭后，可以减小后续的一抗或二抗和膜的非特异性结合，降低背景，增强信噪比。本产品中的Western洗涤液(10X)可以用于Western时一抗或二抗孵育后的洗涤。适当的洗涤可以降低背景，增强信噪比。本产品中的Western二抗稀释液(Secondary Antibody Dilution Buffer for Western Blot) 可以用于Western时二抗(secondary antibody)的稀释。其主要有效成分为经过反复优化的适量BSA、适当的去垢剂和二抗稳定试剂等，一方面能有效减少二抗的非特异性结合，另一方面能有效提升稀释后二抗的稳定保存时间。经本Western二抗稀释液稀释的二抗可以在1-2周内重复使用3-5次。按照每次Western blot需要封闭液10ml、一抗稀释液10ml、二抗稀释液10ml、洗涤液10ml+30ml+30ml+10ml=80ml，约100ml计算，一个包装的本试剂盒，大约可以进行10次Western Blot检测。 注意事项 ： 取放PVDF膜和NC膜应使用平头镊子，并仅轻轻夹取其边角，操作过程须避免膜表面产生划痕、折痕或压痕等痕迹。PVDF膜一经浸润和活化，需一直保持湿润，根据Western进行到的具体步骤可放置于western转模液或洗涤液等适当溶液中，否则可能会产生难以封闭的异常背景。为了能使抗体可以反复多次使用，建议尽量在4℃进行一抗和蛋白膜的结合反应，以减缓一抗的衰减速度。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4℃封闭过夜。本Western洗涤液(10X)配制成Western洗涤液(1X)后，宜4℃保存，并在1-2个月内用完，如果室温保存，宜在1-2周内用完。本Western二抗稀释液经过滤除菌处理，使用过程中应尽量避免细菌污染。一旦启用后建议在一个月内用完，如果发现有沉淀或细菌污染请弃用。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.Western一抗稀释液a.参考所用的一抗的说明，以及样品中目的蛋白的含量，按照适当比例例如1：1000、1：500等比例稀释一抗。一抗稀释后即可直接用于Western。一次Western结束后，可以回收稀释的一抗，4℃保存，以用于下次的Western。2.Western封闭液a.完成转膜后，用Western洗涤液洗涤蛋白酶1-2分钟。加入Western封闭液，封闭60分钟，然后进行一抗孵育等后续操作。3.Western洗涤液(10X)a.取一瓶Western洗涤液(10X)，倒入到一洁净的量筒中，加入蒸馏水混匀并定容到1升后即为Western洗涤液(1X)，并可以用于Western的洗涤。没有用完的洗涤液可以室温保存，通常1-2周内是可以正常使用的。如果发现洗涤液出现浑浊、沉淀等异常现象，宜废弃。b.在一抗或二抗孵育结束后，加入少量Western洗涤液(1X)，使之能盖住蛋白膜，在摇床上洗涤5分钟后，吸尽洗涤液，再加入新的洗涤液洗涤。共洗涤3次，每次5分钟。然后进行后续的操作。如果按照上述洗涤条件Western结果背景仍然较高，可以适当延长洗涤时间，并增加洗涤次数。通常，延长洗涤时间或增加洗涤次数不会对Western结果产生负面影响。4.Western二抗稀释液a.参考所用的二抗的使用说明，以及一抗的质量和样品中目的蛋白的含量，按照适当比例例如1：1000、1：500等比例稀释二抗。二抗稀释后即可直接用于Western。一次Western结束后，可以回收稀释的二抗，4℃保存，以用于下次的Western，通常在1-2周内可以重复使用3-5次。b.详细的Western操作可以参考我们相关网页：http://www.aladdin-e.com/support/western.htm Aladdin's Western Buffer set includes Blocking Buffer, Wash Buffer, Primary Antibody Dilution Buffer, and Secondary Antibody Dilution Buffer commonly used for Western Blot.Primary Antibody Dilution Buffer for Western Blot included in this product can be used for dilution and preparation of primary antibodies. Its main ingredients including BSA, detergent and primary antibody stabilization reagent, have been optimized to reduce non-specific bindings and stabilize primary antibodies after dilution. The diluted primary antibodies in this product can be stored at 4℃ for six months at least, and can be reused multiple times.Blocking Buffer for Western Blot provided in this product can be used to block PVDF or NC membranes. The main active ingredients including BSA and detergent can effectively block the protein binding sites on membranes and reduce non-specific bindings of antibodies, thus reducing the background and enhancing the signal-to-noise ratio.The Western Wash Buffer (10X) can be used to wash membranes after incubation with antibodies. Proper washing can reduce the background and enhance the signal-to-noise ratio.Secondary Antibody Dilution Buffer for Western Blot can be used to dilute secondary antibodies for Western blot. The main active ingredients including BSA, detergent and secondary antibody stabilization reagent, have been optimized for effective reduction of non-specific bindings and stabilization of secondary antibodies after dilution. The diluted secondary antibodies can be reused 3-5 times within 1-2 weeks when stored at 4℃. Precautions ： The PVDF and NC membranes should be handled with flat tip tweezers, and only the corners should be gently gripped, avoiding scratches, creases or indentations of the membrane.The activated PVDF membrane needs to be kept moist in the transfer buffer or wash buffer until use. Once accidentally dried, it needs to be re-activated.To recycle antibodies for later use, we recommend incubating membranes in primary antibody solutions at 4℃ if possible to maintain the activity of primary antibodies.Usually, blocking for 60 minutes at room temperature is sufficient. For some antibodies producing high background, block the membrane overnight at 4℃.The Western Wash Buffer (1X) should be stored at 4℃ and used up within 1-2 months, or within 1-2 weeks if stored at room temperature.Secondary Antibody Dilution Buffer for Western Blot is sterilized by filtration, and bacterial contamination should be avoided during use. At room temperature, it should be used up within one month and discarded once contaminated with microorganisms.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation. Instructions for Use ： 1. Primary Antibody Dilution Buffer for Western Blota. Dilute the primary antibody appropriately according to manufacture's instructions. The diluted primary antibody in this Primary Antibody Dilution Buffer can be recovered and stored at 4℃ for later use.2. Blocking Buffer for Western Blot a. After protein transfer, wash membranes with the Western Wash Buffer for 1-2 minutes, block membranes for 60 minutes in the Blocking Buffer, then incubate in diluted primary antibody solutions.3. Western Wash Buffer (10X) a. Prepare an appropriate amount of Western Wash Buffer (1X) by diluting the Western Wash Buffer (10X) with distilled water and mix well before use. The diluted Wash Buffer can be stored at room temperature for up to 1-2 weeks, and should be discarded when turned turbid or precipitation occurs.b. After incubation of membranes with primary or secondary antibodies, add sufficient Wash Buffer (1X) to fully cover membranes, and wash for 5 minutes with shaking. Repeat washing for a total of 3 times. Then proceed to the subsequent procedures. If there is high background on membrane, extend the washing time or increase the number of washes appropriately, which usually does not have negative impacts on Western results.4. Secondary Antibody Dilution Buffer for Western Blot a. Dilute the secondary antibody appropriately according to manufacture's instructions. The diluted secondary antibody can be recovered and stored at 4℃ for later use. It can be reused 3-5 times within 1-2 weeks.b.

No numbering system information yet

No physical and chemical property information yet

No safety information yet

No production method and use information yet

No upstream information yet

No downstream information yet

No related product information yet