Use of a cyanine dye as a probe for albumin and collagen in the extracellular matrix.
The aim of this work was to develop a quick method for analysis of macromolecules of the extracellular matrix. Of great interest are soluble components of the extracellular matrix, in particular, carrier proteins, whose variation dynamics can characterize the studied tissue in its development, adult stage, and aging. We suggest the method of analysis of the extracellular matrix to reveal the presence of albumin and collagen by using an anionic cyanine dye as a spectral and fluorescence probe. The method was applied for the analysis of the human vitreous body in the course of its development. Albumin was detected by the appearance of the trans monomer absorption and fluorescence bands in the dye spectra, and collagen was detected by the absorption and fluorescence bands of J aggregates. Hyaluronic acid present in the vitreous body does not interfere with the results of the analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis confirmed the presence of albumin in the vitreous body. We suppose that albumin as a protein carrying biologically active macromolecules plays an important role in the processes of differentiation and functional establishment of ocular tissues in the course of their prenatal development.
Panova
Ina G
IG
Kol'tsov Institute of Developmental Biology, Russian Academy of Sciences, ul. Vavilova 26, 119991 Moscow, Russia. pinag@mail.ru
Sharova
Natalia P
NP
Dmitrieva
Svetlana B
SB
Poltavtseva
Rimma A
RA
Sukhikh
Gennady T
GT
Tatikolov
Alexander S
AS
D000418:Albumins / Q000032:analysis*; D001622:Betaine / Q000031:analogs & derivatives* / Q000737:chemistry; D015153:Blotting, Western; D002232:Carbocyanines / Q000737:chemistry*; D003094:Collagen / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D004798:Enzymes / Q000737:chemistry; D005109:Extracellular Matrix / Q000737:chemistry*; D005123:Eye / Q000254:growth & development; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014822:Vitreous Body / Q000737:chemistry
C518775:3,3'-di(gamma-sulfopropyl)-4,5,4',5'-dibenzo-9-ethylthiacarbocyanine betaine pyridinium salt; D000418:Albumins; D002232:Carbocyanines; D004798:Enzymes; D005456:Fluorescent Dyes; D001622:Betaine; D003094:Collagen