JaLCDOI 10.18926/AMO/31741
FullText URL fulltext.pdf
Author Yao, Kenzabroh| Ubuka, Toshihiko|
Abstract <p>A new acidic ninhydrin method for determining free sialic acids is described. The method is based on the reaction of sialic acids with Gaitonde's acid ninhydrin reagent 2 which yields a stable color with an absorption maximum at 470 nm. The standard curve is linear in the range of 5 to 500 nmol of N-acetylneuraminic acid per 0.9 ml of reaction mixture. The reaction was specific only for sialic acids among the various sugars and sugar derivatives examined. Some interference of this method by cysteine, cystine and tryptophan was noted, although their absorption maxima differed from that of sialic acids. The interference by these amino acids was eliminated with the use of a small column of cation-exchange resin. The acidic ninhydrin method provides a simple and rapid method for the determination of free sialic acids in biological materials.</p>
Keywords sialic acid determination acidic ninhydrin reaction acidic ninhydrin method
Amo Type Article
Published Date 1987-12
Publication Title Acta Medica Okayama
Volume volume41
Issue issue6
Publisher Okayama University Medical School
Start Page 237
End Page 241
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 3439478
Web of Sience KeyUT A1987L530300001
JaLCDOI 10.18926/AMO/30439
FullText URL fulltext.pdf
Author Yao, Kenzabroh| Ubuka, Toshihiko| Masuoka, Noriyoshi| Kinuta, Masahiro| Ohta, Jun| Ishino, Kazushi|
Abstract <p>A new method for staining sialoglycoproteins in polyacrylamide gel after disc electrophoresis is described. The method utilizes the reaction of sialic acids with an acidic ninhydrin reagent which yields a stable color with an absorption maximum at 470 nm. After electrophoresis, the polyacrylamide gel is placed in a test tube and heated with 5 ml of the acidic ninhydrin reagent for 10 min in a boiling water bath. Sialoglycoproteins are detected as brown bands. No additional procedure such as destaining is necessary. When 20 micrograms fetuin, a sialoglycoprotein, per gel is applied, the band remains visible for at least 2 h. Stained gel can be scanned with a gel scanner at 470 nm. When the stained gel was dried on a sheet of polypropylene filter, the color was stable for at least one month. The present method is superior to the method using Stains-all (3,3'-diethyl-9-methyl-4,5,4',5'-dibenzothiacarbocyanine) in specificity and simplicity for the detection of sialoglycoproteins.</p>
Keywords sialoglycoprotein polyacrylamide gel electrophoresis staining acidic ninhydrin reaction
Amo Type Article
Published Date 1990-04
Publication Title Acta Medica Okayama
Volume volume44
Issue issue2
Publisher Okayama University Medical School
Start Page 65
End Page 70
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 1694626
Web of Science KeyUT A1990DE10000002