3
$\begingroup$

The molar extinction coefficient of pure antibody is $1.4$ absorbance units per milligram antibody at $\pu{280 nm}$. You have a concentrated antibody solution, so you make a $5$-fold dilution and measure at $\pu{280 nm}$, getting an absorbance of $0.845$. What is the concentration of that solution?

Using Beer's Law this is my work: $$0.845 = (1.4)\cdot (1)\cdot (c) \\ \therefore c = \pu{0.604 mol L^-1}$$

Then I think we need to factor in the dilution, therefore $c = (5) \cdot (0.604) = \pu{3.02 mol L^-1} $. Is this correct?

$\endgroup$
1
  • 2
    $\begingroup$ Your process looks correct to me, but notice that the extinction coefficient is given in AU/mg, so there must be some additional conversion done to get the units correct. $\endgroup$
    – jerepierre
    Commented Oct 2, 2014 at 23:44

1 Answer 1

2
$\begingroup$

Beers law is

$$ E= \epsilon_{\lambda}\ \cdot c\ \cdot d$$

Where $c$ is the concentration of the solution. Then, you've found your answer once you've found $c$.

As Jerepierre points out, if you were to account for units in the answer, the question is actually missing some information. The pathlength $d$ is unaccounted for (but we're assuming its $1$), however if it was say $\mathrm{1\ cm}$, then the units for $\epsilon$ would need to be $\mathrm{AU\cdot mg^{-1} \cdot cm^{-1}}$. Which would mean that the units for your concentration would be in $\mathrm{mg}$. But, for concentration we would need to know antibody per some amount of volume. So taking units into account shows that something is missing.

What you're adding at the end is a step you would take to find the original concentration of the undiluted solution. Given that they diluted the concentration 5 times, but we don't know to what factor so we would have trouble solving for that.

$\endgroup$

Your Answer

By clicking “Post Your Answer”, you agree to our terms of service and acknowledge you have read our privacy policy.

Not the answer you're looking for? Browse other questions tagged or ask your own question.