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Protocol
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Incubation of chemiluminescent or near-infrared Western blots
As an example protocol we would mention the following:
I. Required Reagents
- Blotted nitrocellulose (LI-COR, P/N 926-31090/926-31092) or Immobilon®-FL PVDF membrane (LI-COR, P/N 926-31099 or 926-31100)
- Odyssey® Blocking Buffer (PBS) (LI-COR, P/N 927-40000, 927-40100) or Odyssey Blocking Buffer (TBS) (LI-COR, P/N 927-50000, 927-50100)**
- Primary antibodies
- IRDye® 800CW, 680RD, or 680LT secondary antibodies (LI-COR)
- Tween® 20
- PBS/TBS**
- Ultrapure water
- Methanol for wetting of PVDF
- SDS
** IMPORTANT! PBS-based and TBS-based blocking buffers may be used with this protocol. Odyssey Blocking Buffer is available in both formulations. Be sure to keep your buffer system consistent throughout the protocol for blocking, antibody dilutions, and washes. For example, if you use a TBS-based buffer system, choose Odyssey Blocking Buffer (TBS). If you use a PBS-based buffer system, choose Odyssey Blocking Buffer (PBS).
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