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Useful for research of chondroitin sulfate

Fluoresceinamine labeled Sodium Chondroitin Sulfate

Background

Chondroitin sulfate (CS) is a sulfated glycosaminoglycan composed of repeating disaccharide units of N-acetyl-D-galactosamine (GalNAc) and D-glucuronic acid (GlcUA). CS is abundant in cartilage and exists as unbranched polysaccharide chains covalently linked to the protein core of proteoglycans. These products are prepared by the fluorescent labeling of CS derived from whale cartilage (FACS-A1), shark cartilage (FACS-C1 and D1), squid cartilage (FACS-E1) according to the method of Ogamo et.al.1). Fluoresceinamine molecules are chemically attached to carboxyl groups of the glucuronic acid of CS. These solutions are dissolved in PBS (-) and sterilized by filtration. The endotoxin content is in accordance with the product specifications. 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.


Fluoresceinamine Labeled Sodium Hyaluronate flyer download [PDF]

FACS-A1 FACS-A1 contains approximately one sulfate group per disaccharide unit with about 80% as 4-O-sulfation of GalNAc (A structure unit).
FACS-C1 FACS-C1 contains approximately one sulfate group per disaccharide unit with about 70% as 6-O-sulfation (C structure unit) and about 30% as 4-O-sulfation of GalNAc (A structure unit).
FACS-D1 FACS-D1 contains 20-25 % of disaccharide units with two sulfate groups as 6-O-sulfation of GalNAc and 2-O-sulfation of GlcUA (D structure unit).
FACS-E1 FACS-E1 contains approximately 60 % of disaccharide units with two sulfate groups as 4-O- and 6-O-sulfation of GalNAc (E structure unit).
FACS-P1 FACS-P1 consists of disaccharide units with four sulfate groups as 4-O-, 6-O-sulfation of GalNAc and 2-O-, 3-O-sulfation of GlcUA.


Constitutional formula of Sodium Chondroitin Sulfate


Analysis of binding activities with bFGF

Basic fibroblast growth factor (bFGF, 10 µg/mL in PBS, 100 µL/well) was added into 96well plate (Nunc, 437111) and incubated at 37°C for 1h. The plate was washed with PBS, and then Fluoresceinamine-labeled GAG solution with various concentrations in PBS was added to each well (100 µL/well ) and incubated at 37°C 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.

Analysis of binding activities with BMP-4 by fluorescence correlation spectroscopy (FCS)

Fluorescence correlation spectroscopy (FCS) is an analytical method of molecular- association or dissociation with one molecular level. The interaction of molecules can be detected as a prolongation of average diffusion time (DT) derived from a fluorescent molecule. The figure shows the effect of recombinant human bone morphogenetic protein-4 (rhBMP-4) on the DT of various fluoresceinamine-labeled GAGs. The result indicates relatively strong interactions between rhBMP-4 and FACS-E1, FACS-P1 or FAHep-N1. (J Cell Physiol, 2008, 217(3):769-777)

Specification

Specification
Appearance yellow green solution
Concentration of FACS 0.90〜1.10mg/mL
Wavelength of Excitation 490〜500 nm
Wavelength of Emission 515〜525 nm
Degree of Substitution* 0.600〜1.100%
Fluorescent Purity on Electrophoresis one band at same mobility with original NaCS
Concentration of Endotoxin <0.25 EU/mL>
Storage Store protected from light at -20°C or below. We recommend storing in aliquots appropriate for anticipated usage.

NaCS : Sodium Chondroitin Sulfate

*Degree of Substitution was expressed as the molar percentage of bound fluoresceinamine groups to disaccharide units.


Excitation and emission spectrum of FACS-A1 (pH10)


Elution profiles of FACS-A1, C1, D1, E1 and P1 on GPC
(Column: TSKgel G4000PWXL (7.8 mmI.D. × 30 cm × 1))


Cellulose acetate membrane electrophoresis of FACS-A1
NaCS-A: Sodium Chondroitin Sulfate A

Raw materials of FAHA, FACS, FADS, FAHep and FAHS
Product Name Raw Material Product Name Raw Material
FAHA-H1, H2 Streptococcus sp. FACS-A1 whale cartilage
FAHA-M1, M2 chicken comb FACS-C1 shark cartilage
FAHA-L1, L2 Streptococcus sp. FACS-D1 shark cartilage
FAHA-S1 Streptococcus sp. FACS-E1 squid cartilage
FAHA-T1 Streptococcus sp. FADS-B1 porcine skin
FAHep-N1 porcine intestine FAHS-P1 porcine kidney

<References>
1. Ogamo A et.al.: Carbohydr.Res., 105, 69 (1982)

Product List

Product Name Cat# Quantity Price

Fluoresceinamine Labeled Sodium Chondroitin Sulfate A (A1)

CSR-FACS-A1 3ML

¥ 66,000
$ 880
€ 660

Fluoresceinamine Labeled Sodium Chondroitin Sulfate (C1)

CSR-FACS-C1 3ML

¥ 60,000
$ 800
€ 600

Fluoresceinamine Labeled Sodium Chondroitin Sulfate D (D1)

CSR-FACS-D1 3ML

¥ 66,000
$ 880
€ 660

Fluoresceinamine Labeled Sodium Chondroitin Sulfate E (E1)

CSR-FACS-E1 3ML

¥ 66,000
$ 880
€ 660

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.