Protein Calibration Curves on BioSep-SEC-S (2)

LC Conditions

Column

BioSep 5 µm SEC-s3000 290 Å, LC Column 300 x 7.8 mm, Ea

Brand

Biosep

Part No

00H-2146-K0

Phase Name

SEC-s3000

Elution Type

Isocratic

Mobile Phase

A: 100mM Sodium Phosphate, 300mM NaCl, pH 7.0

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

100

0

Flow Rate

1.00 mL/min

Temperature

22°C

Detection

UV-Vis Abs.-Variable Wave.(UV) @ (22°C)

Detection Info

Sample Notes

Application Focus: To show the calibration curves for the three different BioSep-SEC phases For this application, calibration curves are generated for the three different BioSep-SEC phases using standard proteins of defined molecular weights and non-denaturing operating conditions (100 mM phosphate, 300 mM NaCl, pH 7.0, 1mL/min). For each media (300 x 7.8 mm column) the elution volume where each protein peak elutes was plotted against the log of the expected molecular weight of each protein. A 3rd order polynomial is used to plot the calibration curve for each phase. Specific applications used to generate curves were: App ID# 18892 for BioSep2000, App ID# 18928 for BioSep3000 and App ID#18936 for BioSep4000. While a polynomial was used to generate calibration curves in this example, one could use a linear plot if one removes the data points at the ends of the molecular weight ranges.