Buffer

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Tirumal Rao
Tirumal Rao's picture
Buffer

Blank purpuss,i am using Ammonium Hydrogen carbonate buffer for light scattering of protein conc .Can anybody explain the reason and why i can use sample buffer?

msvnathan
msvnathan's picture
Dear Thirumal,

Dear Thirumal,
Please explain little more about your work which can help me to try to solve the problem.
thank you

Tirumal Rao
Tirumal Rao's picture
Dear Sir,

Dear Sir,
i am working on biotherapeutic proteins .Estimation of protein concentration by light scattering
i am using blank Ammonium hydrogen carbonate buffer
same sample by Photometric method
i am using blank sample buffer what is the reason ?

Dr. Analytical
Dr. Analytical's picture
Do you mean that you analyze

Do you mean that you analyze the "blank" solution and also analyze your protein solution, which is dissolved in the same buffer?

A "blank" measurement is needed for almost every analytical instrument. The blank will determine if there is any response from the solution that your sample is dissolved in. If there is no response, then all the response from your sample is caused by the added components. If you get a response from your blank, then you should subtract this value from any response produced by the sample.

The blank should be exactly the same solution as your sample, except for the sample components. If it is not, then you measurements may not be accurate.

Shubhangi
Shubhangi's picture
In any spectrophotometric /

In any spectrophotometric / light scattering measurement of a protein, it is essential that things other than molecule of interest should not interfere in the measurement.
For this purpose one should do "blanking" with the solution exactly same as sample but without protein of interest. In your case it will be buffer. The value for protein minus value of blank gives the correct value of protein present.