A Secondary Antibody-Detecting Molecular Weight Marker with Mouse and Rabbit IgG Fc Linear Epitopes for Western Blot AnalysisReportar como inadecuado




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Molecular weight markers that can tolerate denaturing conditions and be auto-detected by secondary antibodies offer great efficacy and convenience for Western Blotting. Here, we describe MandR LE protein markers which contain linear epitopes derived from the heavy chain constant regions of mouse and rabbit immunoglobulin G IgG Fc LE. These markers can be directly recognized and stained by a wide range of anti-mouse and anti-rabbit secondary antibodies. We selected three mouse M1, M2 and M3 linear IgG1 and three rabbit R1, R2 and R3 linear IgG heavy chain epitope candidates based on their respective crystal structures. Western blot analysis indicated that M2 and R2 linear epitopes are effectively recognized by anti-mouse and anti-rabbit secondary antibodies, respectively. We fused the M2 and R2 epitopes MandR LE and incorporated the polypeptide in a range of 15–120 kDa auto-detecting markers MandR LE protein marker. The MandR LE protein marker can be auto-detected by anti-mouse and anti-rabbit IgG secondary antibodies in standard immunoblots. Linear regression analysis of the MandR LE protein marker plotted as gel mobility versus the log of the marker molecular weights revealed good linearity with a correlation coefficient R2 value of 0.9965, indicating that the MandR LE protein marker displays high accuracy for determining protein molecular weights. This accurate, regular and auto-detected MandR LE protein marker may provide a simple, efficient and economical tool for protein analysis.



Autor: Wen-Wei Lin , I-Ju Chen , Ta-Chun Cheng, Yi-Ching Tung, Pei-Yu Chu, Chih-Hung Chuang, Yuan-Chin Hsieh, Chien-Chiao Huang, Yeng-Ts

Fuente: http://plos.srce.hr/



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