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Western Blot | Troubleshooting Forum

10/11/2012
Kristie Nybo, Ph.D.

Is it possible that denaturing or freezing the samples cleaves the protein?


BioTechniques Molecular Biology Forum brings bench scientists together in an online community to ask for and share advice regarding laboratory problems. This week’s highlighted question comes from the Protein Methods forum.

Q: I am performing Western blot to look for a 150kDa membrane channel. I get a reproducible, concentration-dependent band that runs at about 110kDa, but no band at 150kDa. This band vanishes when I pre-incubate, so I'm sure it's the primary antibody rather than the secondary that's reacting. Is it possible that denaturing or freezing the samples cleaves the protein?





A: Try epitope tagging the protein and expressing it in…

A: It could be regulatory splice variant.Try RT-PCR to look for…

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