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Western Blot Membrane Incubation with NP-40?

Western Blot Membrane Incubation with NP-40? - Western Blot Forum

Western Blot Membrane Incubation with NP-40? - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 11-10-2007, 04:25 AM
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Question Western Blot Membrane Incubation with NP-40?



Email question from a user:

When performing a ligand blot analysis, under non-reducing conditions, after western blotting, the nitocellulose membrane is incubated in 3% NP-40 for half hour, followed by blocking in BSA in 0.1% Tween PBS. I have come across many papers where NP40 is used. Can you explain to me why this is done?

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Old 11-10-2007, 09:33 PM
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Default Re: Western Blot Membrane Incubation with NP-40?

Because both NP-40 (Igepal) and Tween-20 are both weak nonionic surfactants used in wash solutions for Westerns, ISH, Dot-blots, ELISA. Along with Triton X; another nonionic surfactant, they dissolve easily in TBS, or PBS solutions.
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Old 11-10-2007, 09:56 PM
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Default Re: Western Blot Membrane Incubation with NP-40?

For more background you may want to read "A Guide to the Properties and Uses of Detergents in Biology and Biochemistry" by Srirama M. Bhari
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