Presentation on theme: "Western Blot encyclopedia."— Presentation transcript:
Western Blot encyclopedia
Content Western Blot Principle Western Blot Procedure Western Blot Troubleshooting Western Blot Applications Western Blot Reagents Signaling Pathway Images
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: (1)separation by size, (2)transfer to a solid support, and (3)marking target protein using a proper primary and secondary antibody to visualize.
Western Blot Procedure For more information about western blot protocol, For detailed info about WB protocols, please visit:
Western Blot Troubleshooting Common Problems Recommended Troubleshooting Unusual or unexpected bands Use a fresh sample which had been kept on ice or alter the antibody; Make sure the transfer sandwich is prepared properly; Change the running buffer. Learn more…Learn more… No bands Make sure to use the right and appropriate concentration of antibody; Increase the conc. of antigen; Buffers should be new and uncontaminated; Lower the washing time. Learn more…Learn more… Faint bands or weak signal Increase the conc. of antigen or antibody; Increase exposure time; Change nonfat dry milk into BSA or decrease the amount of milk. Learn more…Learn more… High background Lower the conc. of antibody; Increase washing time; Decrease exposure time. Learn more…Learn more… Patchy or uneven spots Make sure of no air bubbles during transfer; use a shaker for all incubation; Change blocking buffer. For more common problems in western blot and related troubleshooting, please visit:
Western Blot Applications Western Blot for Lyme Western Blot for HIV Western Blot for Herps Western Blot for Lyme Western Blot for FIV Western Blot for HBV