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| Western Blot Forum Discuss western blotting and immunoblot in the western blot forum. |
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| LOL... thanks for the information. I think I may have added too much secondary antibody. WoW.. do you play that game a lot (in the lab maybe jk ) ? ![]() thanks that tip is a good one |
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| A little too much secondary? If your blot is visibly glowing, you need to dilute your secondary about 100-fold. As a bonus, with more dilute secondary, you'll get better specificity and much more stable signal - which equals more reproducible results. Your exposure to film should take 30 seconds-5 minutes. If it's significantly less than this, you are hurting the reproducibility. |
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| Hi I can suggest 4 different ways that you can try in order to get a better result: 1. Dilute your secondary antibody 2. Put the ECL for less time (let's say 60 sec) 3. Dilute your primary antibody 4. Try using two films together By the way do anyone have any suggestion on which software to use in order to quantified bands ? |
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| black , blot , dark , film , glowing , western |
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