Applications

High MW proteins on BioSep4000 (2) and TSK

Analytes

1IgM
2Thyroglobulin
3IgA
4beta-Amylase
5BSA
6Ribonuclease A
7Uridine

Analyte not found

Details

LC Conditions (App ID: 18898)

Column:

BioSep 5 µm SEC-s4000 500 Å, LC Column 300 x 7.8 mm, Ea

Brand Name:

BioSep-SEC-S

Part No:

00H-2147-K0

Phase Name:

SEC-s4000

Elution Type:

Isocratic

Mobile Phase:

A: 100mM Phosphate buffer pH 6.8

Gradient Profile:

Step No.	Time(min)	%A
1	0	100

Flow Rate:

1 mL/min

Temperature:

22°C

Detection:

UV-Vis Abs.-Variable Wave.(UV) @ 220 nm (22°C)

Sample Note:

Application Focus: To compare and contrast BioSep4000 and a leading competitor large range GFC column (TSKgel SWxl 4000) Using a standard mixture of high molecular weight proteins optimized for the 4000 series separation, the BioSep 4000 and a leading competitor columns (TSKgel SWxl 4000) were compared using standard conditions ( 100 mM phosphate pH 6.8, 1 mL/min). Similar amounts of the standard mixture (Ig-M 900KDa, Thyroglobin 670KDa, B-amylase 200 KDa, BSA 66 KDa, Ribonuclease 14K, and Uridine 0.2 KDa) were injected on each column (300 x 7.8 mm) and chromatograms were overlaid. When comparing peak resolutions between columns certain observations can be made regarding the performance of the two columns. In general, better resolution between most components in the mixture is observed for the BioSep 4000 column versus the competitor despite having a smaller elution window between fully excluded and fully included peaks. While the competitor column appears to have better resolution at the very high end of the optimal window (900KDa +) the BioSep 4000 appears to have better resolution between thyroglobin (670KDa) and B-amylase (200KDa), a critical range for many of the application where the BioSep 4000 is used. While these comments may only be relevant for this standard separation, the results indicate that the BioSep 4000 is a good solution for separating high molecular weight proteins.

Order Items Used in This Application

00H-2147-K0

BioSep 5 µm SEC-s4000 500 Å, LC Column 300 x 7.8 mm, Ea

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