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Download the complete western blot guide.
Multiplexing in fluorescent western blotting allows you to:
For more the benefits of using fluorescence in WB see comparison to chemiluminescence here.
Use the green channel (IRDye® 800) to detect your weakest target and the red channel (IRDye® 680) to detect the strongest target.
IRDye® | Species primary was raised | Secondary antibody |
---|---|---|
IRDye® 800CW | Rabbit | Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) |
Mouse | Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) | |
IRDye ® 680RD | Rabbit | Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) |
Mouse | Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) |
See next the fluorescent western blot protocol used by our lab.