Immunoprecipitation of CDT Complexes Containing Deletion Mutants of CdtA (original) (raw)

2013

Abstract

<div><p>(A) The complexes (10 μg each) were immunoprecipitated with 10 μg anti-myc antibody (myc tag was fused to C-terminus of CdtC) in 150 mM NaCl, 20 mM HEPES (pH 7.5), 0.1% Nonidet P40 substitute, followed by exposure to protein G Sepharose beads, and subjected to nondenaturing SDS-PAGE.</p><p>(B) Nonspecific binding of CDT complexes to protein G Sepharose was not detected under the same experimental conditions. A, CdtA; B, CdtB; C, CdtC; A*, truncated CdtA; A1, holotoxin with CdtA Δ18−56; A2, holotoxin with CdtA Δ18−67; A3, holotoxin with CdtA Δ18−75; WT, wild-type; Ab, only anti-<i>myc</i> antibody, no CDT; IgG (H+L), nonreduced immunoglobulin G; IgG H, immunoglobulin G heavy chain; IgG L, immunoglobulin G light chain; SN, supernatant.</p></div

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