2Western Blot Principle Western Blot Procedure ContentWestern Blot PrincipleWestern Blot ProcedureWestern Blot TroubleshootingWestern Blot ApplicationsWestern Blot ReagentsSignaling Pathway Images
3Western 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.
4Western Blot Procedure For more information about western blot protocol,For detailed info about WB protocols, please visit:
5Western 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.
6Western Blot Applications Western Blot for LymeWestern Blot for HIVWestern Blot for HerpsWestern Blot for LymeWestern Blot for FIVWestern Blot for HBV