![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=1722503681&Signature=Adj12W3Sea8dnLs2QwLjpn93I1lrOlmNfpZ2wOTe3HgFeSBVYP983Vnd~jtlj8mhHU0o2OetJTpGkx4CRpe8DxroAyF0VyAMLnVZirqfTs40UHS5pKzAQzaU3IoaqDhEnxAF9lk6oLJUDAItffOe-Jdzu4r6n9HdDSaS6IrddRarzB-4GLtqnOXV4xcGZlo2FhRggYBE5TzzarnnxSZGDcGuG9HMFbxI2y33ArOEAlvc3LPM~7X0iwHJmzxZBzxFDd~NLwJv1HSQ5F4BZXGw3dEqh0wv4rMzkZLLyPdAO9W7ZI6yzBXAs8mv0Q1PkXneDErh0jfJSTYeCxgAdrYujQ__&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
