基于细胞的红外ELISA
R&D Systems®如今推出了基于细胞的红外ELISA，它能测定完整的、经过固定的细胞中磷酸化蛋白及总蛋白水平，而无需裂解细胞。试剂盒可用于贴壁细胞和悬浮细胞，提供了针对目的蛋白和另一个规范化蛋白的抗体。在微孔板的同一个孔中检测两种蛋白，可将各孔之间由于细胞数量的不同所带来的信号差异规范化。基于细胞的红外ELISA使用的二抗带有不同光谱的近红外荧光基团，这些荧光基团与LI-COR® Odyssey®红外激光成像系统兼容。近红外荧光的稳定性强，且信噪比高。
如需更多信息，请访问我们的网站：www.RnDSystems.com/CellBasedELISA.
IGF-I诱导的Akt磷酸化的检测. MCF-7 human breast cancer cells were treated with increasing concentrations of Recombinant Human IGF-I (Catalog # 291-G1) for 20 minutes. After fixation and permeabilization, phosphorylated Akt (S473) and total Akt levels were determined using the Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Cell-Based Infrared ELISA (Catalog # KCBIR887).(A) Each panel shows the same microplate wells with experiments done in duplicate. The images highlight the levels of phospho-Akt (S473; green), total Akt (red), and the ratio of the two (yellow). (B) Histogram profiles of the levels of phosphorylated Akt (S473), normalized to total Akt protein levels in the same well, were generated by analysis of fluorescence intensities. Values represent the mean ± the range of duplicate determinations. Phosphorylated Akt (S473) and total Akt were also detected by Western blot (inset) using the antibodies supplied in the kit.
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