This affinity-purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 50-75 of Isochrysis galbana PCNA.
This affinity purified antibody has been tested for use in ELISA and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band at ~29 kDa in size corresponding to PCNA by western blotting in the appropriate cell
Western blot using Rocklands Affinity Purified anti-Alga PCNA antibody shows detection of a predominant band at ~29 kDa corresponding to PCNA (lane 2). Peptide competition effectively eliminates specific band reactivity (lane 1). For peptide competition, the antibody was pre-incubated with 100 µg (excess) of the peptide for 2h at 37 C followed by overnight incubation at 4 C with 100 µg (excess) of the peptide, followed by a brief centrifugation and then reaction with the membrane. Each lane contains a lysate from 105 cells from the marine dinoflagellate Prorocentrum minimum. Cells were separated by SDS-PAGE and transferred onto nitrocellulose using standard techniques. After blocking the membrane was probed with the primary antibody diluted to 1:200. Incubation was followed by washes and reaction with HRP Gt-a-Rabbit IgG [H&L] MX (611-103-122). Detection was by ECL using a 30 sec exposure time. Other detection systems will yield similar results. Personnel Communication. S. Lin, U.Conn.
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