Transcript Western Blot PPT.

Western Blot encyclopedia
http://www.western-blot.us
Content
• Western Blot Principle
• Western Blot Procedure
• Western Blot Troubleshooting
• Western Blot Applications
• Western Blot Reagents
• Signaling Pathway Images
http://www.western-blot.us
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.
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Western Blot Procedure
For more information about western blot protocol,
For detailed info about WB protocols, please visit:
http://www.western-blot.us/western-blot-protocol
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Western Blot Troubleshooting
For more common problems in western blot and related troubleshooting, please visit:
http://www.western-blot.us/guide-to-the-problem-of-wb
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…
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…
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…
High background
• Lower the conc. of antibody;
• Increase washing time;
• Decrease exposure time. Learn more…
Patchy or uneven
spots
• Make sure of no air bubbles during transfer;
• use a shaker for all incubation;
• Change blocking buffer.
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Western Blot Applications
Western Blot for Lyme
Western Blot for Lyme
Western Blot for HIV
Western Blot for FIV
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Western Blot for Herps
Western Blot for HBV
Western Blot Reagents
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Signaling Pathways – Free for download by visit
http://www.sinobiological.com/Signaling-Pathways-a-5590.html
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