不是二抗，胜似二抗 —— Mouse IgG Binding Protein[新品推荐]

在繁花似锦，蓬勃发展的生命科学领域里，二抗的存在犹如尘埃，低微到被所有的人忽略。但是，这小小的二抗却是免疫学中不可或缺的必备之物，HRP标记的二抗用来进行WB、IHC，ELISA；FITC/PE等荧光标记的二抗可以用来进行IF、流式FCM。。。传统的二抗制备工艺也相对简单，通常是经过免疫动物制备纯化多抗，再进行各种标记，生物个体之间的差异或多或少就会带来抗体批次间的差异，导致不同批次间抗体的灵敏度和效价的差别。众所周知，Santa Cruz 是世界著名的抗体生产商，提供了数以万计的抗体，包括不断推出的单克隆抗体之外，近期还发布了了一种替代传统抗小鼠二抗的新型“二抗”——Mouse IgG Binding Protein。

Mouse IgG Binding Protein，即小鼠结合蛋白，与小鼠IgG轻链（IgGκ /IgGλ ）特异性结合，可替代传统的山羊/兔抗小鼠二抗，用于各种免疫学实验。相对于传统二抗，小鼠结合蛋白的特异性和灵敏度更高，检测背景更低，而且几乎完全消除了传统方法生产的二抗所具有的批次间差异。Santa Cruz 提供的小鼠结合蛋白标记种类也十分丰富，包括：HRP（fig.1）、FITC （fig.2） 、PE、Biotin、 CruzFluor™ 荧光 (CFL 488/555/594) （fig.3），另外还提供可以在LI-COR Odyssey系统上检测的近红外荧光标记：CruzFluor™ 680 (CFL 680) 和CruzFluor™ 790 (CFL 790) （fig.4） ，可满足传统二抗所能进行的一切实验需求。

接下来，就让我们来体验一下使用 Mouse IgG Binding Protein 带来的精彩绝伦的实验结果吧！

另外，除了满足常规的 Western Blot 之外，针对目前越来越被广大科研者认可的近红外荧光检测，即：Near-Infrared (NIR)  western blot ，Mouse IgG Binding Protein 产品还提供可以在 LI-COR Odyssey系统上检测的近红外荧光标记： CruzFluor™ 680 (CFL 680) 和 CruzFluor™ 790 (CFL 790) ，可满足用户更加精准的蛋白质表达定量分析。

Figure 4 : Simultaneous infrared western blot analysis of β-Actin expression, detected with β-Actin (C4): sc-47778 and m-IgGκ BP-CFL 680: sc-516180 and HSP 70/HSC 70 expression, detected with HSP 70/HSC 70 (W27): sc-24 and m-IgGλ BP-CFL 790: sc-516195 in HeLa (A), DU 145 (B), MCF7 (C), MDA-MB-231 (D), HCT-116 (E) and HUV-EC-C (F) whole cell lysates.

来自客户的试用反馈，也显示了使用 Mouse IgG Binding Protein替代传统山羊/兔抗小鼠二抗可使 western blot 检测背景更低（fig.5），灵敏度更高（fig.6）。

Works great, will no longer use R anti-mouse HRP!!

I received a small free vial of this anti-mouse-HRP alternative with one of SCBT order and was a bit apprehensive about using it without a side by side comparison with a conventional anti-mouse-HRP from a vendor that I have been using for more than 10 years. I was blown away with by the results. Attached is the side by side comparison of anti-mouse-HRP (the 2 left blots) vs. the kappa BP-HRP using the same lysates and same primary antibodies. This is far superior to anti-mouse-HRP secondary and I have been using this exclusively since this first set of blots.

Very nice on blots

We compared our standard goat anti-mouse IgG-HRP with this mouse kappa binding protein-HRP conjugate side by side on blots with identical samples and the same primary antibody, sc-365744 mouse monoclonal anti-Na,K-ATPase alpha3 H-4. At a 1:2,000 dilution of both the secondary antibody and the anti-kappa-BP HRP, the kappa BP gave a much stronger signal. See the 1 sec exposure in the image. There seems to be a lot of confusion among users about what this probe actually is. Many refer to it as an antibody, but it isn't, it's a binding protein. Whether it is Protein L or something else is not clear from the current datasheet. It would be useful to see evidence for species specificity. For this purpose, though, it is very good.

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