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Control variation

  • OldForum
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13 years 3 months ago #1 by OldForum
Control variation was created by OldForum
I am performing an inhibition assay. I generally run my control several times throughout the experiment to make sure that the chip is regenerating/functioning properly. I have noticed that over time, I get a decrease in signal when running the control. I thought perhaps the chip was no good, however when I prepare fresh control, it works as well as the initial preparation. My control is an antibody (I have a peptide coupled to BSA coupled to the chip). Could this reflect antibody instability at low concentration? Its only a few hours difference? I am totally baffled. I thought at first it might be a regen issue but that does not seem to be the case. Any thoughts?
rhaspel

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  • OldForum
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13 years 3 months ago #2 by OldForum
Replied by OldForum on topic Control variation
It is possible that the antibody is sticking to the vial, decreasing the concentration and the response. Esspecially at low concentration this can be noticable.

You can try to coat the vials with BSA but then you can get some bffer jumps due to BSA detaching the vial.
You can try some non sticky vials or coat them self with repel silane.
Arnoud

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