Immobilon^® Block - PO (Phosphoprotein Blocker)

The possible causes and potential remedies for challenges encountered as a result of blocking, washing, antibody incubation, and detection/exposure of Western blots

蛋白磷酸化是一种可逆的翻译后加工，在细胞内信号传递中有非常重要的功能。采用Western Blotting技术检测磷酸化蛋白是探讨信号传导中上游调控、下游功能调整、信息交流及反馈机制等的基本手段。我们提供专用的优质试剂和研究方法，帮助你获得准确、灵敏的磷酸化检测数据。

我们明白。西部片的污点可能会让人头疼，我们当然也有过这样的经历。因此，请与我们分享您的丑陋污点，并获得一份惊喜！虽然我们不能告诉你是什么--那会破坏惊喜的气氛--但它会激发实验室的创造力。

We get it. Westerns can be a pain in the blot, and we’ve certainly made our share. So, share with us your ugly blots and receive a surprise! And while we can’t tell you what it is – that would spoil the surprise - except that it will spark creativity in the lab.

Antibody reuse for Western blotting is a common practice for many researchers. While many antibodies lose potency with time or degrade even faster due to improper storage conditions, it is important to recognize the potential value of recovering the primary antibody for possible reuse in some experiments. The SNAP i.d.® 2.0 system is not only able to reduce the immunodetection processing time, but its flexibility lets you combine conditions used in the standard immunodetection protocol and also allows the collection of antibody for future reuse. Here, we compare the antibody recovery and reuse in the standard immunodetection protocol with the antibody recovery and reuse in SNAP i.d.® system using the extended protocol and the original SNAP i.d.® protocol.

In Western blotting, blocking of unbound membrane sites is necessary to prevent non-specific binding of the antibodies which otherwise would lead to a high background on the blot. The traditional milk blocker can work well; however, inadvertent variations in its composition can lead to irreproducible results. Such variations include the milk’s concentration, fat content, solubility, detergent quality, and numerous other factors. Bløk reagents are a family of protein-free, noisecancelling reagents that reduce background for consistent, quality results. They are available in three room temperature-stable, ready-to-use formulations for Chemiluminescence and chromogenic detection, Fluorescence detection, and Phosphoprotein immunodetection. The protein-free nature of Bløk reagents allows for membranes to be stained with chromogenic reagents, such as Ponceau S or Coomassie™ blue, after blocking or immunodetection. Bløk reagents have been tested and validated for Western blot, dot blot, and ELISA applications.