Question

We did a lab on Gel Filtration Chromatography in which we used two "standards" Phenol Red (included in gel) and Blue Dextran(excluded from gel). An unknown protein (either hemoglobin or myoglobin) was given. Question is why is a standard used (in relation to the lab) and how is it useful? Please provide a detailed answer. Thank You.

Answer 1

Gel Chromatography based on elution volum principal. In which molecule flow through gel and its pore so larger moleculer weight Blue Dextran will flow only side by beads not entering in to pores so excluded quickly and time will be measured void volume equal to elution volume

On the other hand phenol red low molecular weight copound will flow thorough pores of gel it will take more time so elution volume will be more than void volume

Based on staderdization of above two unknown protein hemoglobin or myoglobin elution volume vs void volume determined