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Western Blot encyclopedia.

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Presentation on theme: "Western Blot encyclopedia."— Presentation transcript:

1 Western Blot encyclopedia

2 Content Western Blot Principle Western Blot Procedure Western Blot Troubleshooting Western Blot Applications Western Blot Reagents Signaling 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: (1)separation by size, (2)transfer to a solid support, and (3)marking 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 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:

6 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

7 Western Blot Reagents

8 Signaling Pathways – Free for download by visit


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