25 ug of the indicated Human whole cell extracts was loaded onto a 12% SDS-PAGE gel. 3 ug/mL of the antibody was used in this experiment. The canonical 41 kDa protein is shown, as well as isoforms at 37 and 20 kDa that also contain the peptide sequence (Isoforms 1, 2 & 3).
Surface Plasmon Resonance Kinetic Characterization of Polyclonal Antibody Affinity. Purified polyclonal antibodies were immobilized on a Protein A/G coated Carterra LSA sensor chip (PAGH200M) at concentrations of 5, and 50 ug/mL in duplicate. Antibodies on the surface were exposed to interaction with peptides sequentially via microfluidic controlled flow at 333nM peptide concentration for kinetic characterization of the binders for affinity and specificity, followed by curve fitting using the Kinetics software. Kd determinations for both concentrations were averaged and results and standard deviation are shown.
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