Characterizing TRAIL-Heparin Binding

To characterize the binding of WT TRAIL and TRAIL mutants to heparin, 100 μg of purified WT or mutant TRAIL was applied to a 1 ml HiTrap heparin–Sepharose column (Cytiva Lifesciences) and eluted with a salt gradient from 150 mM to 1 M NaCl at pH 7.1 in 25 mM HEPES buffer. The conductivity measurements at the peak of the elution were converted to the concentration of NaCl based on a standard curve.

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Luo Y., Hao H., Wang Z., Ong C.Y., Dutcher R., Xu Y., Liu J., Pedersen L.C, & Xu D. (2024). Heparan sulfate promotes TRAIL-induced tumor cell apoptosis. eLife, 12, RP90192.

Publication 2024

HiTrap heparin–Sepharose column

Corresponding Organization : National Institutes of Health

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Variable analysis

independent variables

WT TRAIL
TRAIL mutants

dependent variables

Binding of WT TRAIL and TRAIL mutants to heparin

control variables

Applied 100 μg of purified WT or mutant TRAIL to a 1 ml HiTrap heparin–Sepharose column
Eluted with a salt gradient from 150 mM to 1 M NaCl at pH 7.1 in 25 mM HEPES buffer
Conductivity measurements at the peak of the elution were converted to the concentration of NaCl based on a standard curve

controls

No positive or negative controls were explicitly mentioned in the provided information.

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