Skip to Content
MilliporeSigma

HPLC Analysis of Protein Standards by Size Exclusion on Zenix® as Affected by Pore Size: 100Å vs. 300Å

HPLC Analysis of Protein Standards by Size Exclusion on Zenix® as Affected by Pore Size: 100Å vs. 300Å application for HPLC

Materials

analytical column

Product No.
Description
Pricing

Zenix® SEC-100 Gel Filtration Column, 3μm

L × I.D. 30 cm × 4.6 mm, 3 μm particle size, stainless steel

Zenix® SEC-300 Gel Filtration Column, 3 μm

L × I.D. 15 cm × 4.6 mm, 3 μm particle size, stainless steel

Zenix® SEC-300 Gel Filtration Column, 3 μm

L × I.D. 30 cm × 4.6 mm, 3 μm particle size, stainless steel

standard

Product No.
Description
Pricing

Myoglobin from equine heart

≥90% (SDS-PAGE), essentially salt-free, lyophilized powder

Uracil

≥99.0%

Thyroglobulin from bovine thyroid

(For Gel Filtration Chromatography)

IgG from rabbit serum

reagent grade, ≥95% (SDS-PAGE), essentially salt-free, lyophilized powder

Bovine Serum Albumin

lyophilized powder, ≥96% (agarose gel electrophoresis)

CONDITIONS

column

Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 μm particles (Z777028)

column

Zenix SEC-100, 30 cm x 4.6 mm I.D., 3 μm particles (Z777006)

column

Zenix SEC-300, 15 cm x 4.6 mm I.D., 3 μm particles (Z777026)

column

Zenix SEC-100, 15 cm x 4.6 mm I.D., 3 μm particles

mobile phase

0.15 M sodium phosphate, monobasic, pH 7.0 with sodium hydroxide

flow rate

0.3 mL/min

pressure

1215 psi (84 bar)

column temp.

25 °C

detector

UV, 215 nm

injection

0.5 μL

sample

Waters BEH125 SEC protein standard

Description

Analysis Note

With size exclusion chromatography, pore size fundamentally determines the relative retention of analytes. Larger pores permit inclusion of larger analytes, smaller pores exlcude larger analytes. This application compares the retention and selectivity of protein standards on columns of widely differing pore sizes as well as the sequential combination of both. The combination of the two in series provides an intermediate selectivity. Each 30 cm column was compared to combination of the two 15 cm columns connected in series; in the case of the latter, the SEC-300 preceding the SEC-100 column in the flow path.

Legal Information

Zenix is a registered trademark of Sepax Technologies