Heparan sulfate (HS) is a sulfated glycosaminoglycan composed of repeating disaccharide units of D-iduronic acid (IdoUA) or D-glucuronic acid (GlcUA) and N-acetyl-D-glucosamine or N-sulfo-glucosamine. HS is abundant in basement membrane of kidney and blood vessel exists as unbranched polysaccharide chains. This product is prepared by the fluorescent labeling of HS derived from porcine kidney according to the method of Ogamo et al.1). Fluoresceinamine molecules are chemically attached to carboxyl groups of the GlcUA or IdoUA of HS. This solution is dissolved in PBS (-) and sterilized by filtration. The excitation wavelength is 490~500 nm and the emission wavelength is 515~525 nm. The enclosed Certification of Analysis lists actual values for product specifications.
Typical disaccharide formula of Sodium Heparan Sulfate
Analysis of binding activities with bFGF
Basic fibroblast growth factor (bFGF, 10μg/mL in PBS, 100 μL/well) was added into 96well plate (Nunc, 437111) and incubated at 37℃ for 1h. The plate was washed with PBS, and then fluorescein-labeled GAG solution with various concentrations in PBS was added to each well (100μL/well ) and incubated at 37℃ for 1h. After washing the plate, 0.1N NaOH was added (100μL/well), and the fluorescence intensity was measured using a fluoro-plate reader with an excitation wavelength of 485 nm and an emission wavelength of 535 nm. It was observed that the fluorescence intensity was increased with the concentration of FACS-E1, FAHS-P1 FAHep-N1 and FAHS-P1 whereas the intensity was little increased in the case of FAHA-M1. The result suggests an interaction between sulfated GAG and bFGF.
