Does buffer composition affect the hillock coefficient contained by binding assays?
Also, what about the IC50 value of a drug?
I need a hint if possible.
Answers:
Normally, I would so no. Sometimes, however, there are situations where the presence of Na+ for example, will affect the bearing a ligand binds to a receptor, but the actual composition of the buffer shouldn't. So, e.g. if you use Tris buffer vs. HEPES buffer, you should see the same Hill number, and then if you add Na+ to any buffer, for the particular case I referred to, you would see the effect of this cation in both buffers. Same for IC50 for a drug. It shouldn't rise and fall with alterations in buffer composition, and besides, one usually compares IC50 values to others, and they should all be determined using alike conditions. An IC50 by itself is not of much use.
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I need a hint if possible.
Answers:
Normally, I would so no. Sometimes, however, there are situations where the presence of Na+ for example, will affect the bearing a ligand binds to a receptor, but the actual composition of the buffer shouldn't. So, e.g. if you use Tris buffer vs. HEPES buffer, you should see the same Hill number, and then if you add Na+ to any buffer, for the particular case I referred to, you would see the effect of this cation in both buffers. Same for IC50 for a drug. It shouldn't rise and fall with alterations in buffer composition, and besides, one usually compares IC50 values to others, and they should all be determined using alike conditions. An IC50 by itself is not of much use.
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