Characterization of alpha-1-antitrypsin by isoelectric focusing on an ultrathin polyacrylamide gel layer

1979 ◽  
Vol 53 (1) ◽  
pp. 91-95 ◽  
Author(s):  
G. Massi ◽  
A. Fabiano ◽  
D. Ragusa ◽  
P. Auconi
Keyword(s):  
Isoelectric Focusing ◽  
Polyacrylamide Gel ◽  
Gel Layer ◽  
Alpha 1 Antitrypsin

Concanavalin A-horseradish peroxidase bridge staining of alpha-1 glycoproteins separated by isoelectric focusing on polyacrylamide gel.

1976 ◽  
Vol 24 (8) ◽  
pp. 908-914 ◽  
Author(s):  
R C Allen ◽  
S S Spicer ◽  
D Zehr
Keyword(s):  
Horseradish Peroxidase ◽  
Isoelectric Focusing ◽  
Concanavalin A ◽  
Periodic Acid ◽  
Polyacrylamide Gel ◽  
New Method ◽  
Polyacrylamide Gels ◽  
Periodic Acid Schiff ◽  
Coomassie Blue

The Coomassie Blue protein stain and the periodic acid-Schiff stain for glycoproteins are compared to a new method of staining glycoproteins resolved electrophoretically. The method utilizes a Concanavalin A-horseradish peroxidase sequence to visualize selectively glycoproteins with terminal or internal mannose or terminal N-acetylglucosamine. The method applied to characterization of M and Z allele products of alpha-l-antitrypsins separated by isoelectric focusing of polyacrylamide gels slabs have revealed differences in carbohydrate content of various components that were previously undetected.

Isolation and characterization of goat serum α1-globulin protease inhibitors

1982 ◽  
Vol 60 (1) ◽  
pp. 28-35 ◽  
Author(s):  
Albert Hercz
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Protease Inhibitors ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Isolation And Characterization ◽  
Sds Page

α1-Globulin-type protease inhibitors were isolated from goat serum by two methods, namely preparative isoelectric focusing and preparative electrophoresis in polyacrylamide gel. The fractions obtained by the first method showed varying isoprotein compositions by analytical isoelectric focusing. Sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE) revealed the presence of one protein in the fractions with the same velocity of migration as purified human α1-antitrypsin and a second protein with a slightly higher migration velocity. The ratios of trypsin-inhibiting to chymotrypsin-inhibiting capacities in all the fractions were the same and both inhibitors were stable upon storage. The reaction of the inhibitors with trypsin and chymotrypsin was also demonstrated by analytical isoelectric focusing.The fractions obtained by preparative gel electrophoresis (the second method) contained the same proteins but their proportions varied widely in different fractions as demonstrated by analytical electrofocusing in the presence of urea and by SDS–PAGE. The early fractions, which consisted predominantly of α1-antitrypsin, showed a high inhibiting capacity for trypsin and none or only negligible capacity for chymotrypsin. Conversely, in the late fractions, the proportions of the proteins and inhibiting capacities were reversed. At 4 °C the trypsin-inhibiting capacity was stable for weeks but the chymotrypsin-inhibiting capacity of the preparation rapidly decreased.These observations indicate that the inhibition of proteases by goat α1-globulins is due to at least two closely associated but distinguishable proteins. One of these, corresponding to human α1-antitrypsin, would have an appreciable capacity to inhibit trypsin, but unlike the latter, little or no capacity for chymotrypsin inhibition. The inhibition of chymotrypsin is due to the second, unidentified α1-globulin.

scholarly journals The rapid purification and partial characterization of human serum angiotensinogen

1987 ◽  
Vol 243 (1) ◽  
pp. 121-126 ◽  
Author(s):  
C J Campbell ◽  
P A Charlton ◽  
C J Grinham ◽  
C J Mooney ◽  
J E Pendlebury
Keyword(s):  
Sialic Acid ◽  
Human Serum ◽  
Isoelectric Focusing ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Multiple Forms ◽  
Rapid Purification ◽  
Blue Sepharose

Human angiotensinogen has been purified 390-fold from serum by a rapid high-yielding procedure that involved chromatography on Blue Sepharose, phenyl-Sepharose, hydroxyapatite and immobilized 5-hydroxytryptamine (5-HT). Angiotensinogen was specifically bound to immobilized 5-HT, which effected a partial resolution into multiple forms, which were also evident when analysed by SDS/polyacrylamide-gel electrophoresis (Mr 59,400, 60,600, 62,600 and 63,800). This heterogeneity was confirmed by resolution into six main bands on isoelectric focusing, ranging from pI 4.40 to 4.82. N-terminal analysis, digestion with human renal renin and deglycosylation studies implied that the preparation comprised several forms of angiotensinogen, varying in their degree of glycosylation. The presence of sialic acid was shown to be a major factor in determining the heterogeneity.

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