murtas100

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2024-11-27 22:31:29

Sudden problems with western-blots

Let’s gather some more info. 😁

1732748217
We are using Bio Rad Turbo Transfer System, which I belive is semi-dry, in which there is in-build turbo mode, which take 3 minutes. And yeah, our next guess, is that we're using too much secondaries, but we will check it no faster than monday. This low dilution of secondaries was established experimentally before I joined the project, and was so low I'm guessing most likely due to the age of antibodies? Perhaps the new ones require stronger dilutions for good results (manufacturer does propose 1:10 000/1:20 000 dilution).

1732753319
I didn’t see mention, but what are you using for your chemiluminescence reagent? They often have guidelines as well.

1732822864
We use Clarity Western ECL Substrate, also made by BioRad. In general, we use a lot of their equipment.

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