title: A Secondary Antibody-Detecting Molecular Weight Marker with Mouse and Rabbit IgG Fc Linear Epitopes for Western Blot Analysis.
abstract: 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 M&R 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 (M&R LE) and incorporated the polypeptide in a range of 15-120 kDa auto-detecting markers (M&R LE protein marker). The M&R LE protein marker can be auto-detected by anti-mouse and anti-rabbit IgG secondary antibodies in standard immunoblots. Linear regression analysis of the M&R 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 M&R LE protein marker displays high accuracy for determining protein molecular weights. This accurate, regular and auto-detected M&R LE protein marker may provide a simple, efficient and economical tool for protein analysis.
authors: Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan.
authors: Wen-Wei
authors: WW
authors: Lin
authors: Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: I-Ju
authors: IJ
authors: Chen
authors: Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Ta-Chun
authors: TC
authors: Cheng
authors: Department of Public Health and Environmental Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Yi-Ching
authors: YC
authors: Tung
authors: Department of Medical Laboratory Science and Biotechnology, College of Health Sciences, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Pei-Yu
authors: PY
authors: Chu
authors: Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Chih-Hung
authors: CH
authors: Chuang
authors: Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Yuan-Chin
authors: YC
authors: Hsieh
authors: Graduate Institute of Clinical Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.|Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Chien-Chiao
authors: CC
authors: Huang
authors: Department of Biochemistry, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Yeng-Tseng
authors: YT
authors: Wang
authors: Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Chien-Han
authors: CH
authors: Kao
authors: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
authors: Steve R
authors: SR
authors: Roffler
authors: Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan.|Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.|Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, Kaohsiung, Taiwan.|Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors: Tian-Lu
authors: TL
authors: Cheng
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western / Q000379:methods*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D007074:Immunoglobulin G / Q000235:genetics; D051379:Mice; D008970:Molecular Weight; D011817:Rabbits
chemical_list: D000906:Antibodies; D015415:Biomarkers; D000939:Epitopes; D007074:Immunoglobulin G