A Quantitative Method for Detecting Ara h 2 by Generation and Utilization of Monoclonal Antibodies

Figure 5

Characterization of the specificity of mAbs by Western blotting. (a) Isotyping of the IgG1 heavy chain of mAbs. Lane 1: 1-2E10-IgG1. Lane 2: 2-1D5-IgG2a. Lane 3: 3-1C5-IgG1. Lane 4: 4-1C2-IgG1. Lane 5: 5-1G4-IgG1. (b) Characterization of the specificity of the Ara h 2.02 mAbs. Lane 1: protein marker; lane 2: peanut extract before desalting; lane 3: rAra h 2; 1-2E10, 2-1D5, 3-1C5, and 4-1C2 were the mAbs applied individually. (c) Lane 1: peanut extract before desalting; lane 2: peanut extract after desalting; M: prestained protein marker; Ctrl: nonimmune serum; Serum: immunized serum; 3-1C5: the mAb was applied; CBB: Coomassie brilliant blue staining. (d) Lane 1: rAra h 2; lane 2: peanut extract after desalting; 1-2E10, 2-1D5, 3-1C5, 4-1C2, and 5-1G4 were the mAbs applied individually. (e) Lane 1: roasted peanut extract; lane 2: raw peanut extract; M: prestained protein marker; 1-2E10, 2-1D5, 3-1C5, and serum (immune serum) were applied individually. CBB: Coomassie brilliant blue staining.

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