What concentration of antibody should I use for an antibody that has never been tested in immunoblotting?
• The antibody will require optimization testing just as it would for traditional western blot testing. We recommend starting at a high concentration. Refer to supplier’s recommended concentration for western blotting, then use 3- to 5 times the concentration in 1/3 to 1/5 the volume for SNAP i.d. immunodetection. Q. What is the best approach to optimize an antibody for use with the SNAP i.d. System? • By using two triple well blot holders, six different concentrations of primary antibody can be evaluated in parallel. See application note entitled: “Triple Well Blot Holder as a Tool for 30-Minute Western Blotting Optimization with the SNAP i.d. Protein Detection System” (AN1965EN00). Q. If I have to leave the lab for longer than 10 minutes during the incubation period, will my experiment be damaged? • No, you can leave the SNAP i.d. unattended for periods longer than 10 minutes and the membrane will not dry out or affect the immunodetection. Antibody incubation periods of 30 minutes to
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- What concentration of antibody should I use for an antibody that has never been tested in immunoblotting?
