UltraBio™ Western检测增敏剂

W751599-100ml

100ml

阿拉丁生产的UltraBio™ Western检测增敏剂(UltraBio™ Western Blot Enhancer)是一种通过直接用于一抗或二抗的稀释，增强Western检测时抗原抗体间的特异性结合，从而可以使Western检测的灵敏度增强数倍至十几倍的增敏试剂。 本产品通过大规模精细筛选获得很可能通过粘附、缠绕或捆绑特异性结合的抗原抗体复合物的大分子化合物，以及配套的降低背景的封闭试剂和去垢剂组合，从而有效解决Western检测过程中经常遇到的低灵敏度和高背景等问题，极大地提高了信噪比。本产品也可用于免疫荧光(Immunofluorescence)、免疫组化(Immunohistochemistry)、免疫斑点印迹(Dot Blot)和ELISA等其它涉及抗原抗体结合的免疫检测的增敏，也可以尝试用于其他蛋白相互作用检测的增敏。 Western，也称Western blot、Western blotting、Western印迹，常简写为WB，中文称为免疫印迹，是用抗体检测蛋白表达的主要方法之一。通常的步骤是SDS-PAGE电泳、转膜、封闭、一抗孵育、二抗孵育和二抗标记物检测，可参考阿拉丁的网页详细介绍：http://www.aladdin-e.com/。 Western blot实验过程中，会遇到如下的常见问题：目的蛋白丰度低或抗体效价低而导致目的蛋白条带信号弱；封闭效果不好而导致背景高；抗体特异性不好而导致非特异性条带强等问题。解决此类问题通常需要经过长时间的条件优化，特别是更换抗体或增加抗体用量会增加实验成本。 本产品可极大提高目的条带的信号强度：本产品通过促进抗原抗体的特异性结合，产生的信号强度会比传统方法高数倍甚至十几倍，可以提高低丰度蛋白的检测成功率或节省蛋白样品的用量。 本产品使获得的检测结果更佳：本产品可有效降低背景，并且使目的条带信号更强，容易获得更理想的Western检测结果。 本产品可提高抗体特异性：本产品中包含了高效的封闭试剂和降低非特异性结合的去垢剂组合，可以有效提高抗体的特异性，降低非特异性条带的信号，获得更好的Western检测结果。 本产品可节省抗体使用量：与传统的方法相比，本产品的检测信号会增强很多，可以适当减少一抗和二抗的用量，有效节约实验成本。同时，本产品稀释和配制的一抗或二抗可以在4℃保存，后续可以反复多次用于Western，利于节约一抗和二抗。 本产品使用便捷：本产品是一种即用型溶液，直接用于一抗和二抗的稀释配制即可，无须区分一抗稀释用或二抗稀释用，不增加任何额外的实验步骤，不改变常规的Western检测步骤。对于较难检测的目的蛋白，可以节省用于目的蛋白检测优化的时间和精力。 本产品的印迹膜兼容性好：本产品兼容PVDF (Polyvinylidene Fluoride, 聚偏二氟乙烯)膜和NC (Nitrocellulose, 硝酸纤维素)膜。 本产品的检测底物兼容性好：本产品的组分对各种标记抗体的酶活性均没有影响，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，适用于化学发光法(即ECL法)、比色法和荧光法检测。 本产品应用范围广：本产品特别适用于Western检测，也可用于免疫荧光、免疫组化、免疫斑点印迹和ELISA等其它涉及抗原抗体相互作用的免疫检测。 本产品的性能达到甚至超过了国际同类产品。本产品(UltraBio™ Enhancer)与普通抗体稀释液(without enhancer)及知名的国外M公司的同类产品(Competitor M)的Western检测效果对比参见图1。仅使用不同的一抗和二抗的稀释试剂，其余实验条件完全一致。结果显示，与普通抗体稀释液(without enhancer)相比，使用本产品获得的目的条带信号显著增强；与国外M公司的同类产品相比，在目的条带信号基本一致的情况下，使用本产品的非特异性条带信号显著减弱。 图1. UltraBio™ Western检测增敏剂与普通抗体稀释液(without enhancer)及国外M公司的同类产品(Competitor M)的Western 检测效果对比。蛋白样品为HeLa细胞裂解液，每组从左到右蛋白的上样量依次为2.5μg、5μg和10μg。转膜完成后，使用阿拉丁的UltraBio™ Western封闭液封闭10分钟，用图中注明的不同的试剂配制的一抗(NF-κB p65 Rabbit Polyclonal Antibody, AF0246, 1:5000)溶液室温孵育1小时，洗涤，然后使用图中注明的不同的试剂配制的二抗(辣根过氧化物酶标记山羊抗兔IgG(H+L), A0208, 1:3000)溶液室温孵育1小时，洗涤后，ECL Star 孵育，然后曝光相同时间成像。实际结果会因蛋白样品、抗体、检测仪器等的不同而存在差异，图中数据仅供参考。 本产品与Merck公司的 Immunoreaction Enhancer Kit、Thermo公司的 Western Blot Enhancer属于同类产品。 注意事项 ： 为了确保本产品最佳的检测效果，推荐配套使用阿拉丁生产的P0252 QuickBlock? Western封闭液或P0023B Western封闭液，和P0023C Western洗涤液。 本产品在低温保存或-20℃冻存溶解后很可能会有少量白色沉淀产生，此时可以在30-50℃水浴中适当孵育一会，并在沉淀完全溶解后使用。 本产品稀释和配制的一抗或二抗使用后可以在4℃保存，后续可以反复多次用于Western检测。稀释配制好的一抗或二抗使用次数较多后会出现检测效果下降的情况，如果出现检测效果下降、浑浊、沉淀等情况宜丢弃。 为了能使稀释的一抗或二抗可以反复多次使用，抗体孵育结束后稀释的抗体应立即存放在4℃，以便于后续重复使用。 本产品推荐用于一抗或二抗的稀释，不建议用于替代印迹膜的封闭，也不建议用于ELISA检测中抗原或抗体的包被(coating)。 本产品经过优化本身具有很好的封闭活性，用于一抗、二抗的稀释时无须再添加其它的封闭试剂如BSA、脱脂奶粉、酪蛋白等。如果额外添加其它封闭试剂，可能会产生一定的负面效果。 由于目的蛋白和抗体的不确定性，本产品有可能对某些目的蛋白和相应抗体的检测信号强度的提高效果一般。 本产品实际使用过程中，如果目的条带信号非常强的情况下，非特异性条带的信号也会相应地有所增强。如果非特异性条带距离目的蛋白条带很近，影响检测效果，可以尝试适当降低蛋白上样量、降低一抗二抗浓度或缩短曝光时间。 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 ： 1.参考所用抗体的说明，以及样品中目的蛋白的含量，使用本产品按照适当比例稀释一抗或二抗。抗体稀释后即可直接用于Western或其它检测，Western或其它检测的方法步骤不变。Western等检测结束后，可以回收稀释的抗体，4℃保存，以用于下次的Western等检测。2.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。 Aladdin's UltraBio™ Western Blot Enhancer provides a simple method to improve the detection sensitivity of Western Blot by promoting the specific binding of antigens with antibodies. This product can be used to directly dilute primary and secondary antibodies and enables an increased detection sensitivity by several to more than ten times.The UltraBio™ Western Blot Enhancer consisted of a large molecular compound that binds with antigen-antibody complex, background-reducing blocking reagent, and detergent, greatly improving the signal-to-noise ratio of Western Blot by effective reduction of membrane background and improvement of detection capacity. This product can also be used to sensitize other antigen-antibody binding involved immunoassays, such as Immunofluorescence, Immunohistochemistry, Immuno-dot Blot, and ELISA. The application of this product to sensitize the detection of protein-protein interactions can also be tested.During the Western blot, the following problems are often encountered, including weak signal of target protein due to its low abundance or the low titer of its antibody, high background caused by poor blocking, and non-specificity resulting from the poor specificity of antibody. It is time-consuming and costly to improve Western results using traditional methods. However, with the UltraBio™ Western Blot Enhancer, all these problems can be solved simply and effectively.This product greatly improves the signal intensity of the target protein by promoting the specific binding of antigen with antibody. Compared to the traditional method, this product increases the signal intensity by several to more than ten times, which is particularly helpful in detecting target proteins with low-abundance or rare samples.This product increases the signal-to-noise ratio by reducing background and improving antibody specificity. It contains a high-efficient blocking reagent and detergent that reduces the background and the non-specific binding of antibodies.This product saves antibodies. Compared with traditional methods, the signal intensity of the protein band is enhanced greatly, and therefore, the dosage of primary and secondary antibodies can be appropriately reduced. In addition, primary and secondary antibodies diluted in this product can be recovered and stored at 4ºC to be reused multiple times.This product is easy to use Precautions ： To obtain the best performance, we recommend using this product in combination with QuickBlock™ Western Blocking Buffer (, P0252) or Western Blocking Buffer (, P0023B), and Western Washing Buffer (, P0023C).A small amount of white precipitates may occur in this product after stored at low temperatures or frozen at -20°C. Before use, the precipitates should be completely dissolved by incubating the bottle in a 30-50°C water bath for an appropriate period of time.Store the diluted antibodies at 4℃ in the SignalUp™ Western Blot Enhancer. The diluted antibodies can be recovered and reused multiple times. Discard when the diluted antibodies demonstrate decreased sensitivity with turbidity or precipitation.To enable the diluted antibodies to be reused as many times as possible, the antibody dilutions should be stored at 4ºC immediately after each use.This product is recommended for diluting antibodies, not for blocking Western membranes, nor coating antigens and antibodies for ELISA detection.This product has been optimized to have good blocking performance and can be used directly for antibody dilution. Some negative effects may occur if additional blocking reagents are added.Due to the uncertainties of target protein and antibody, the performance of this product might be modest in improving the signal intensity of certain target proteins.SignalUp™ Western Blot Enhancer improves the intensity of both target protein and non-specific proteins. When the visualization of the target protein is affected by the neighboring non-specific proteins, less protein loading, higher antibody dilution, or reducing the exposure time can be tested.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. Dilute primary or secondary antibodies with an appropriate amount of SignalUp™ Western Blot Enhancer, according to antibody instructions and the content of target proteins in the sample. All other Western Blot procedures should be followed exactly. The diluted antibody can be recovered and stored at 4ºC for future uses.2. For detailed Western Blot protocol, please visit our website at: http://www.aladdin-e.com/support/western.htm