Take the 2-minute tour ×
Biology Stack Exchange is a question and answer site for biology researchers, academics, and students. It's 100% free, no registration required.

I have a few questions:

  1. What is an OD value?

  2. Why do we use blank solution in Lowry's protein estimation method?

  3. If The OD of a protein is 0.01, what does it mean?

Thanks in advance.

share|improve this question

1 Answer 1

  1. The OD measurement is the output of what the photometer measures. It is actually the amount of light which is scattered or absorbed by your sample - scientifically called extinction.
  2. A blank is used to be able to substract the influence of reagents, light that is scattered on the surfaces of the cuvette (which is probably also not completely clean) and so on. Every of this influences reduces the amount of light which should reach the detector on the other side of the probe and give you false extinction readings. If you are using a colored detection sample (which colors deeper or looses color during the reaction) this is more obvious. The solution already has an extinction on its own which is not of interest. So you make a blank (which basically substracts all these influences from you sample reading) to correct for this. How this is done depends on the photometer. Some make a correction reading at the beginning, while others use a second cuvette (a blank cuvette) which is permanently measured.
  3. It means that the protein gives you this extinction. Without a calibration curve (made from serial dilutions with known protein concentrations) it is impossible to make any further comment on this number.
share|improve this answer
    
Thanks a lot for quick response.but can you elaborate ans no. 2 please. –  N00B Apr 28 at 16:43
    
@N00B Sure, I expanded my answer. –  Chris Apr 28 at 18:15
2  
You can think of the blank as a way of calibrating the instrument. The blank defines the amount of light that will hit the photodetector when there is none of the species of interest in the cuvette. By setting this as the zero you have prepared the instrument for measuring the samples of interest. –  Alan Boyd Apr 28 at 18:18

Your Answer

 
discard

By posting your answer, you agree to the privacy policy and terms of service.

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