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Ligan Binding Calculations

# Ligan Binding Calculations - Step 2 A=cl With an extinction...

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. Step 2 =ℰ A cl , With an extinction coefficient of 33 900 M - 1 ·cm - 1 , , and a path length of 1cm we . should get an absorbance of 0 2 if the chromophore is in a concentration given by = / c A = l . / * = . 0 2 33900 1 5 9 x 10 - 6 M so we are looking for our bound HABA concentrations to be in a range around . 6μM . Step 4 . We want the absorbance of about 0 2 to be near the point where half the , HABA is bound to avidin so since the binding of avidin with its ligands is very , . tight the avidin should be about twice the concentration of HABA We saw in . step 2 that this is about 6μM So we want all the tubes to have about 12 μM . ; / . avidin Avidin is expensive we only have about 1mL of 25mg mL of material , The MW of the avidin tetramer is 68 kD and there must be at least 4 . / receptors per tetramer So since the 25mg mL stock solution is the same as 25 / ( ), ( / )/( g L just changing the units we can write 25g L 68x10 3 / ) = . g 4 mol 1 47mM is . the minimum concentration of receptor in the avidin stock solution ,

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Ligan Binding Calculations - Step 2 A=cl With an extinction...

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