![Fig. 2. Binding of HS oligosaccharides to MIP-1α. / An MIP-1α (BB10010) affinity gel column was prepared by mixing 100 μg MIP-1α with 100 μg heparin in 500 μL coupling buffer (0.1 M HEPES [N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid], 80 mM NaCl, pH 7.0) and was incubated for 20 minutes at room temperature. MIP-1α was then bound to Affi-Gel 10, and the column was prepared as previously described.14 A control column was prepared with the MIP-1α omitted. 3H-labeled HS chains were digested with platelet heparanase under different pH conditions, and the resultant HS fragments were size-fractionated on a Cl-6B Sepharose gel filtration column. Intact HS and fragments of 20 kDa,14 kDa, 10 kDa, and 5 kDa were applied to the MIP-1α (BB10010) and to a control Affi-Gel column in 0.15 M NaCl and were eluted with a stepwise NaCl gradient. The percentage of material eluting at more than 0.15 M NaCl has been averaged from 2 experiments, after subtraction of the control results. Binding of the different oligosaccharides to MIP-1α is expressed as a percentage of the binding of the intact parent HS molecule. Standard error bars are shown. Data are from the experiments described in Stringer et al.10](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/101/6/10.1182_blood-2002-08-2588/4/h80633985002.jpeg?Expires=1735500674&Signature=KM7ntJ7i182n9LpSFiSwXWM4NDXQjPbLUtUym0TEiE~G4~uSJopgm0v3bp4it8mnLjBsgUrwbJQPloNkePJeJsUu5JWwsiS20TMSf6mrTsOSYEkd48QDPoLvA9tkNF9Z7NQhniKYqWjK8jRlAkTJlD7iXg2UhHKa9XNuNxQ7V5KWfd1Rhc~7flQB2VqdXRVhFoogGp~tkXeVMwVZMyrTXa11AH6DJnzwf1b1CQ7NwayDNaWlwvodxDE6iKRn2doeJbjg4WYTwadQuuYZCjMARKSHc9IT7vsnXKCnCeAWDB-4RvakQietbFC8cYQmlrsWQO9mBl7k48s-mhZa~eLCYg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Binding of HS oligosaccharides to MIP-1α.
An MIP-1α (BB10010) affinity gel column was prepared by mixing 100 μg MIP-1α with 100 μg heparin in 500 μL coupling buffer (0.1 M HEPES [N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid], 80 mM NaCl, pH 7.0) and was incubated for 20 minutes at room temperature. MIP-1α was then bound to Affi-Gel 10, and the column was prepared as previously described.14 A control column was prepared with the MIP-1α omitted. 3H-labeled HS chains were digested with platelet heparanase under different pH conditions, and the resultant HS fragments were size-fractionated on a Cl-6B Sepharose gel filtration column. Intact HS and fragments of 20 kDa,14 kDa, 10 kDa, and 5 kDa were applied to the MIP-1α (BB10010) and to a control Affi-Gel column in 0.15 M NaCl and were eluted with a stepwise NaCl gradient. The percentage of material eluting at more than 0.15 M NaCl has been averaged from 2 experiments, after subtraction of the control results. Binding of the different oligosaccharides to MIP-1α is expressed as a percentage of the binding of the intact parent HS molecule. Standard error bars are shown. Data are from the experiments described in Stringer et al.10
