UltraBio™封闭液(10X)

阿拉丁生产的UltraBio™封闭液(UltraBio™ Blocking Buffer)是最新一代的快速高效封闭液，总体效果显著优于传统的基于BSA(牛血清白蛋白)、脱脂奶粉、酪蛋白(Casein)等的封闭液及国外同类产品，可以用于Western blot (WB)、Immunofluorescence (IF)、Immunohistochemistry (IHC)、Immunocytochemitry (IC)等实验中的封闭、一抗或二抗的稀释。本产品为10倍浓缩(10X)溶液，不含去垢剂。UltraBio™封闭液快速高效。封闭时间通常仅需 5-15分钟，并且和BSA、脱脂奶粉、酪蛋白等传统封闭液以及国外同类的快速封闭液相比，显示出更强的信噪比(参考图1)。UltraBio™封闭液封闭后背景极低。本封闭液不含血清和白蛋白，确保极高的信噪比。UltraBio™封闭液兼容性好，兼容辣根过氧化物酶(Horseradish peroxidase, HRP)、碱性磷酸酶和生物素标记的二抗。本产品中添加了不影响HRP和AP活性的防腐剂，不会干扰HRP或AP标记二抗的检测。同时本产品不含生物素，不会干扰基于生物素的检测。UltraBio™封闭液(10X)使用灵活。本产品为 10X溶液，可以根据实验需要使用PBS、TBS或其它适当溶液稀释至1X，即可用于相关实验。同时也可以根据实验需要自行添加Tween-20或Triton X-100至终浓度为0.05%-0.1%后用于封闭、一抗或二抗的稀释。本产品与BSA及国外同类产品的封闭效果对比参见图1。在相同样品和实验条件下，仅封闭液及封闭时间存在如下图所示的差异时，阿拉丁的UltraBio™封闭液封闭后的整体背景明显低于BSA封闭后的背景，而且目的条带亮度明显高于国外同类品牌产品。 图1. UltraBio™封闭液与BSA及国外同类产品的封闭效果比较。每组实验从左到右依次为：5 μl蛋白Marker，2.5 μg蛋白量的HeLa细胞裂解液，5μg蛋白量的HeLa细胞裂解液。请注意对于Antibody A，UltraBio™封闭液比BSA和国外同类产品具有更低的背景，并且和国外同类产品相比信号明显更强。对于Antibody B，UltraBio™封闭液比BSA具有更低的背景，和国外同类产品相比信号明显更强。实际实验结果会因样品、抗体、实验条件等的不同而存在差异，图中数据仅供参考。关于不同封闭液的比较和选择，请参考阿拉丁的相关网页：http://www.aladdin-e.com/。按照每张膜封闭需要5-10ml UltraBio™封闭液(1X)计算，一个包装的本产品可以封闭100-200张膜。 注意事项 ： 本产品推荐仅使用一次，重复使用可能会导致封闭效果下降。但对于一些信噪比很高的一抗，例如一些内参抗体，本封闭液可以重复使用2-3次。回收的封闭液请勿与未使用过的封闭液混合。通常本产品用于PVDF膜及NC膜时的封闭时间为5-15分钟。对于一些背景非常高的抗体，可以尝试将封闭时间延长为30-60分钟。此外，如有特殊需要，也完全可以 4℃封闭过夜。由于没有任何一种封闭液是适用于所有实验体系的，因此对于一些特殊的实验，可能需要根据具体情况考虑使用其它更合适的封闭液。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转膜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。 在背景较高的情况下，为进一步提高信噪比，本产品稀释至1X后可以自行添加Tween-20或Triton X-100至终浓度为0.05%-0.1%。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. QuickBlock™封闭液(10X) 的稀释根据实验需要，使用PBS、TBS或含有0.05%-0.1% Tween-20或Triton X-100的PBS或TBS稀释 QuickBlock™封闭液(10X)至1X。2. Western Blot中膜的封闭a.完成转膜后，用Western洗涤液洗涤蛋白膜1-2分钟。b.根据膜的大小，在平皿或者其它适当容器中倒入一定体积的QuickBlock™封闭液(1X)，确保封闭液后续能充分覆盖膜即可。对于常规的western，一张约6.6×8.5cm的膜推荐使用约10ml左右的封闭液。c.用平头镊夹住膜的一角，将膜放置在QuickBlock™封闭液(1X)中，使封闭液完全浸没膜，置于水平摇床上封闭约10分钟(通常5-15分钟均可；经多种抗体的测试封闭10分钟的效果很多时候会显著优于常规的BSA封闭1小时的效果)。d.封闭后的膜即可用于一抗孵育等后续实验。详细的Western操作可以参考如下的相关网页:http://www.aladdin-e.com/support/western.htm。3. IF、IHC等实验的封闭按照相关实验步骤，直接用QuickBlock™封闭液(1X)替换传统封闭液即可，一般封闭时间可以缩短至 10分钟(在阿拉丁测试过的多个一抗中，封闭10-20分钟的效果无明显差别，封闭10分钟的效果等同于或显著优于常规的封闭方法)。 UltraBio™ Blocking Buffer from Aladdin is a new generation of rapid and highly effective blocking buffer. Its overall effect is significantly better than that of traditional blocking buffers based on BSA (bovine serum albumin), skim milk powder, casein and the like, as well as similar products from abroad. It can be used for blocking in Western blot (WB), immunofluorescence (IF), immunohistochemistry (IHC), immunocytochemitry (IC) and other experiments, primary or secondary antibody dilution. This product is a 10-fold concentrated (10X) solution that does not contain a detergent. UltraBio™ blocking solution is fast and efficient. The blocking time is usually only 5-15 minutes, and compared with traditional blocking solutions such as BSA, skim milk powder, casein, and similar fast blocking solutions from abroad, it shows a stronger signal-to-noise ratio (see Figure 1). UltraBio™ blocking solution has a very low background after blocking. The blocking solution does not contain serum or albumin, ensuring an extremely high signal-to-noise ratio. UltraBio™ blocking solution is compatible with horseradish peroxidase (HRP), alkaline phosphatase and biotin-labeled secondary antibodies. This product contains a preservative that does not affect the activity of HRP and AP, and will not interfere with the detection of HRP- or AP-labeled secondary antibodies. It also does not contain biotin, which will not interfere with biotin-based detection. UltraBio™ Blocking Solution (10X) is flexible to use. This product is a 10X solution that can be diluted to 1X with PBS, TBS or other appropriate solution according to experimental needs and then used in related experiments. It can also be used for blocking, primary antibody or secondary antibody dilution by adding Tween-20 or Triton X-100 to a final concentration of 0.05%-0.1% according to experimental needs. The blocking effect of this product compared with BSA and similar foreign products is shown in Figure 1. Under the same sample and experimental conditions, only when there are differences in the blocking solution and blocking time as shown in the figure below, the overall background after blocking with UltraBio™ blocking solution from Aladdin is significantly lower than that after blocking with BSA, and the brightness of the target band is significantly higher than that of similar foreign brands. Figure 1. Comparison of the blocking effect of UltraBio™ blocking solution with BSA and similar foreign products. From left to right in each group of experiments: 5 μl protein marker, 2.5 μg protein amount of HeLa cell lysate, 5 μg protein amount of HeLa cell lysate. Please note that for Antibody A, UltraBio™ blocking solution has a lower background than BSA and similar foreign products, and the signal is significantly stronger than similar foreign products. For Antibody B, UltraBio™ Blocking Buffer has lower background than BSA and a significantly stronger signal than foreign similar products. Actual experimental results may vary due to differences in samples, antibodies, experimental conditions, etc. The data in the figure is for reference only. For a comparison and selection of different blocking solutions, please refer to the relevant Aladdin webpage: http://www.aladdin-e.com/. Based on the calculation that 5-10 ml of UltraBio™ blocking solution (1X) is required to block each membrane, one package of this product can block 100-200 membranes. Note: This product is recommended for single use only. Reuse may result in a decrease in blocking efficiency. However, for some primary antibodies with a high signal-to-noise ratio, such as some internal reference antibodies, this blocking solution can be reused 2-3 times. Do not mix the recovered blocking solution with unused blocking solution. Usually, this product is used for PVDF membranes and NC membranes with a blocking time of 5-15 minutes. For some antibodies with very high backgrounds, you can try to extend the blocking time to 30-60 minutes. In addition, if there is a special need, it is completely fine to block overnight at 4 °C. Since no blocking solution is suitable for all experimental systems, for some special experiments, it may be necessary to consider using other more suitable blocking solutions based on specific circumstances. Flat-tipped forceps should be used to pick up and place the PVDF membrane and NC membrane, and only the edges should be gently clamped. During the operation, scratches, creases or indentations on the membrane surface should be avoided. Once the PVDF membrane is wetted and activated, it needs to be kept wet at all times. According to the specific steps of Western, it can be placed in an appropriate solution such as a western transfer solution or washing solution. Otherwise, an abnormal background that is difficult to seal may be generated. In the case of high background, to further improve the signal-to-noise ratio, this product can be diluted to 1X and then Tween-20 or Triton X-100 can be added to a final concentration of 0.05%-0.1%. This product is for professional scientific research use only, and is not intended for clinical diagnosis or treatment, nor for use in food or drugs. For your safety and health, please wear laboratory clothing and disposable gloves when handling. Instructions for use: 1. Dilution of UltraBio™ Blocking Buffer (10X) Dilute UltraBio™ Blocking Buffer (10X) to 1X with PBS, TBS or PBS or TBS containing 0.05%-0.1% Tween-20 or Triton X-100 according to the needs of the experiment. 2. Blocking of membranes in Western Blot a. After the transfer is complete, wash the protein membrane with Western washing solution for 1-2 minutes. b. Depending on the size of the membrane, pour a certain volume of UltraBio™ blocking solution (1X) into a petri dish or other appropriate container to ensure that the blocking solution can fully cover the membrane later. For a conventional western, it is recommended to use about 10 ml of blocking solution for a membrane about 6.6 × 8.5 cm. c. Use flat-tipped forceps to hold one corner of the membrane, place the membrane in UltraBio™ blocking solution (1X), completely submerge the membrane in the solution, and place it on a horizontal shaker to block for about 10 minutes (usually 5-15 minutes is fine; testing with multiple antibodies has shown that the effect of 10 minutes of blocking is often significantly better than the effect of the conventional 1-hour BSA blocking). d. The blocked membrane can then be used for subsequent experiments such as primary antibody incubation. 3. For IF, IHC and other experiments, simply follow the relevant experimental procedures and directly replace the traditional blocking solution with UltraBio™ blocking solution (1X). The general blocking time can be shortened to 10 minutes (among the multiple primary antibodies tested by Aladdin, there is no significant difference in the effect of blocking for 10-20 minutes, and the effect of blocking for 10 minutes is equivalent to or significantly better than the conventional blocking method).

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