Simultaneous detection of submicrogram quantities of hyaluronic acid and dermatan sulfate on agarose-gel by sequential staining with toluidine blue and Stains-All

2005 ◽  
Vol 820 (1) ◽  
pp. 131-135 ◽  
Author(s):  
N VOLPI ◽  
F MACCARI ◽  
J TITZE
Keyword(s):  
Hyaluronic Acid ◽  
Toluidine Blue ◽  
Dermatan Sulfate ◽  
Simultaneous Detection ◽  
Agarose Gel

Heparin and sulfated mucopolysaccharides—a micro system for quantitative differentiation and determination

1977 ◽  
Vol 55 (5) ◽  
pp. 1179-1189 ◽  
Author(s):  
Louis B. Jaques ◽  
Tak K. Sue ◽  
Norman M. McDuffie ◽  
Sandra M. Wice
Keyword(s):  
Aqueous Solution ◽  
Hyaluronic Acid ◽  
Optical Density ◽  
Toluidine Blue ◽  
Linear Relation ◽  
Cetylpyridinium Chloride ◽  
Agarose Gel ◽  
Azure A ◽  
Tissue Extracts ◽  
The Individual

Heparin (Hep), hyaluronic acid, chondroitins (sulfate) A, B, and C, and heparitins (sulfate) A, B, C, and D were subjected to microelectrophoresis in barbital–agarose gel, fixed with cetylpyridinium chloride and stained with toluidine blue. The optical densities of the resulting bands were compared with optical densities obtained upon reaction with azure A in aqueous solution and with the carbazole reagent. A linear relation was obtained between optical density and concentration of purified sulfated mucopolysaccharide (SMP). Less than 1 μg of Hep and 2 μg of other SMPs are required for measurement by electrophoresis, while about 30 μg of each is required with the carbazole reagent. The optical density of a mixture of SMPs was equal to the sum of the densities for the individual SMPs upon microelectrophoresis. It was demonstrated that the individual SMPs in mixtures were distinguished by reaction with specific enzymes and by changes in migration in agarose with barbital, phthalate, ethylenediamine, or propane-diamine buffers, permitting ready demonstration and quantitation of various SMP species. Examples are shown of the application of the procedure to measure the total SMPs and individual SMPs in tissue extracts. The method is sensitive, reproducible, flexible, and measures quantities [Formula: see text] of those measured colorimetrically, yet is relatively unaffected by protein, carbohydrate, or inorganic electrolytes.

Glycosaminoglycan biosynthesis in the isolated perfused rat lung

1984 ◽  
Vol 57 (6) ◽  
pp. 1648-1654 ◽  
Author(s):  
P. M. Sampson ◽  
R. B. Boyd ◽  
G. G. Pietra ◽  
A. P. Fishman
Keyword(s):  
Hyaluronic Acid ◽  
Heparan Sulfate ◽  
Dermatan Sulfate ◽  
Rat Lung ◽  
Electron Microscopic ◽  
Chondroitin Sulfates ◽  
Isolated Lung ◽  
Microscopic Studies

The suitability of an isolated lung, perfused under carefully monitored conditions, for the study of the biosynthesis of glycosaminoglycans was examined for the rat lung using either [35S]-sulfate or [6-3H]glucosamine. Metabolic and electron-microscopic studies after 3 h of perfusion showed that under the conditions of this study the isolated lung showed no anatomical or metabolic derangements. All glycosaminoglycans normally synthesized in the intact lung were identified. The predominant glycosaminoglycan was heparan sulfate (40% of total). Approximately 14% of the glucosamine incorporated into the glycosaminoglycans was found in hyaluronic acid. Less than 5% of either label was in heparin. The remainder of the synthesized glycosaminoglycans, with the exception of 10% which could not be identified, consisted of the chondroitin sulfates and dermatan sulfate. The relative proportions of the newly synthesized glycosaminoglycans, including the low amounts of heparin, are similar to those found in isolation of endogenous lung glycosaminoglycans. The isolated perfused rat lung appears to be a useful model for the study of glycosaminoglycan biosynthesis by the intact lung.

Online preconcentration and determination of chondroitin sulfate, dermatan sulfate and hyaluronic acid in biological and cosmetic samples using capillary electrophoresis

2019 ◽  
Vol 42 (17) ◽  
pp. 2867-2874 ◽  
Author(s):  
Kanokporn Chindaphan ◽  
Kanet Wongravee ◽  
Thumnoon Nhujak ◽  
Thasinas Dissayabutra ◽  
Monpichar Srisa‐Art
Keyword(s):  
Capillary Electrophoresis ◽  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Dermatan Sulfate ◽  
Online Preconcentration

Oocytes Preserve the Ability of Mouse Cumulus Cells in Culture to Synthesize Hyaluronic Acid and Dermatan Sulfate

1993 ◽  
Vol 160 (2) ◽  
pp. 405-412 ◽  
Author(s):  
Evelina Tirone ◽  
Gregorio Siracusa ◽  
Vincent C. Hascall ◽  
Gaetano Frajese ◽  
Antonietta Salustri
Keyword(s):  
Hyaluronic Acid ◽  
Dermatan Sulfate ◽  
Cumulus Cells ◽  
Cells In Culture

scholarly journals Localization of hyaluronic acid in human articular cartilage.

1994 ◽  
Vol 42 (4) ◽  
pp. 513-522 ◽  
Author(s):  
A Asari ◽  
S Miyauchi ◽  
S Kuriyama ◽  
A Machida ◽  
K Kohno ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Dermatan Sulfate ◽  
Keratan Sulfate ◽  
Human Articular Cartilage ◽  
Middle Zone ◽  
Deep Zone ◽  
Weak Staining ◽  
Pre Treatment

To demonstrate localization of hyaluronic acid (HA) in articular cartilage of the human femur, biotinylated HA-binding region, which specifically binds HA molecules, was applied to the tissue. In sections fixed by 2% paraformaldehyde-2% glutaraldehyde, HA staining was detected in lamina splendens and chondrocytes in the middle zone. By pretreatment with trypsin, intense HA staining appeared in the extracellular matrix of the deep zone and weak staining in the superficial and middle zones. Moreover, pre-treatment with chondroitinase ABC (CHase ABC) intensely enhanced the stainability for HA in the superficial and middle zones and weakly in the deeper zone. Combined pre-treatment of trypsin with CHase ABC abolished intra- and extracellular staining for HA in all zones. By microbiochemical study, the concentrations of HA and dermatan sulfate were high in the middle zone, whereas those of chondroitin sulfate and keratan sulfate were high in the deep zone. These results suggest that HA is abundantly synthesized in and secreted from the chondrocytes, particularly in the middle zone, whereas it is largely masked by proteoglycan constituents in the extracellular matrix.

Improved Preservation of the Tissue Surrounding Percutaneous Devices by Hyaluronic Acid and Dermatan Sulfate in a Human Skin Explant Model

2009 ◽  
Vol 38 (3) ◽  
pp. 1098-1110 ◽  
Author(s):  
Antonio Peramo ◽  
Cynthia L. Marcelo ◽  
Steven A. Goldstein ◽  
David C. Martin
Keyword(s):  
Hyaluronic Acid ◽  
Human Skin ◽  
Dermatan Sulfate

scholarly journals Production of GJycosaminoglycans by Rat Hair Follicle Cells in Vitro

1981 ◽  
Vol 34 (2) ◽  
pp. 171 ◽  
Author(s):  
R Frater ◽  
D Hewish
Keyword(s):  
Hyaluronic Acid ◽  
Heparan Sulfate ◽  
Hair Follicle ◽  
Culture Medium ◽  
Dermatan Sulfate ◽  
Hair Follicles ◽  
Follicle Cells ◽  
Cell Aggregates ◽  
Dermal Tissue

Cultures of cells from rat dermal tissue, containing a large proportion of cells from the hair follicles, were found to produce glycosaminoglycans. The glycans were associated with the cell aggregates which typically form in such cultures, and also appeared to be present in material which was released from the cells. Examination of the glycosaminoglycan species present in the cultures showed the presence of hyaluronic acid, dermatan sulfate, chondroitin-4-sulfate, heparin and heparan sulfate-C, the first two compounds being secreted.into the culture medium. The patterns of synthesis and sulfation of glycosaminoglycans were found to change with continued time in culture.

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