Anti-cd33 antibodies and method for treatment of acute myeloid leukemia using the same
A technology of antibody and identity, applied in the field of antibody binding to CD33
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[0199] [164] 1.3. Preparation and sequencing of heavy and light chain samples
[0200] [165] The heavy and light chains of the muMy9-6 antibody were separated by SDS-PAGE under reducing conditions. Reduced and denatured antibodies were run on 12% Tris-glycine gels (Novex, San Diego, CA). After electrophoresis, gels were spotted onto Immobilonpsq membranes using CAPS / MeOH buffer. After transfer, membranes were stained with Ponseau S. The bands corresponding to the light and heavy chains were excised for protein sequencing.
[0201] [166] The light chains of the antibodies were sequenced directly from the membrane on an ABI 494 Procise sequencer by automated Edman degradation chemistry.
[0202] [167] The N-terminus of the heavy chain was blocked, so the protein was digested in situ with trypsin according to Gharahdaghi et al. (1996). The digested mixture was then analyzed by MALDI-TOF mass spectrometry on a Kratos Kompact SEQ device. Selected peptides were subjected to MS / ...
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