2 Western Blot Principle Western Blot Procedure ContentWestern Blot PrincipleWestern Blot ProcedureWestern Blot TroubleshootingWestern Blot ApplicationsWestern Blot ReagentsSignaling Pathway Images
3 Western Blot Principle Western blot is an important technique used in cell and molecular biology. By using a western blot, researchers are able to identify specific proteins from a complex mixture of proteins extracted from cells.The basic principle of western blot is to use three elements to accomplish this task:separation by size,transfer to a solid support, andmarking target protein using a proper primary and secondary antibody to visualize.
4 Western Blot Procedure For more information about western blot protocol,For detailed info about WB protocols, please visit:
5 Western Blot Troubleshooting For more common problems in western blot and related troubleshooting, please visit:Common ProblemsRecommended TroubleshootingUnusual or unexpected bandsUse a fresh sample which had been kept on iceor alter the antibody;Make sure the transfer sandwich is preparedproperly;Change the running buffer. Learn more…No bandsMake sure to use the right and appropriateconcentration of antibody;Increase the conc. of antigen;Buffers should be new and uncontaminated;Lower the washing time. Learn more…Faint bands or weak signalIncrease the conc. of antigen or antibody;Increase exposure time;Change nonfat dry milk into BSA or decreasethe amount of milk. Learn more…High backgroundLower the conc. of antibody;Increase washing time;Decrease exposure time. Learn more…Patchy or uneven spotsMake sure of no air bubbles during transfer;use a shaker for all incubation;Change blocking buffer.
6 Western Blot Applications Western Blot for LymeWestern Blot for HIVWestern Blot for HerpsWestern Blot for LymeWestern Blot for FIVWestern Blot for HBV
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