Genetic Variants of Alpha 1-Antitrypsin and Hemoglobin Typed by Isoelectric Focusing in Preselected Narrow pH Gradients with Phastsystem

1992 ◽  
Vol 38 (4) ◽  
pp. 577-580 ◽  
Author(s):  
J O Jeppsson ◽  
R Einarsson
Keyword(s):  
Isoelectric Focusing ◽  
Genetic Variants ◽  
Clinical Medicine ◽  
Ph Gradients ◽  
Total Analysis Time ◽  
Fast Screening ◽  
Abnormal Hemoglobin ◽  
Total Analysis ◽  
Protein Staining ◽  
Alpha 1 Antitrypsin

Abstract In this method for automatically running and staining isoelectric focusing (IEF) gels, pre-made dehydrated polyacrylamide gels were rehydrated before assays run with the PhastSystem (Pharmacia LKB Biotechnology). The typing of genetic variants of hemoglobin and alpha 1-antitrypsin in narrow pH gradients (pH 6.7-7.7 and 4.2-4.9, respectively) was simple, convenient, and reproducible. The clinically important variants of alpha 1-antitrypsin (ZZ and SZ) were identified from serum or dried blood on filter paper. The fast screening of abnormal hemoglobin samples (HbS) for cases in clinical medicine was easily performed. The total analysis time for the phenotyping with conventional protein staining was approximately 60 min.

Isoelectric focusing of serum proteins on modified thin cellulose-acetate membranes.

1979 ◽  
Vol 25 (7) ◽  
pp. 1320-1322 ◽  
Author(s):  
J Ambler ◽  
G Walker
Keyword(s):  
Cellulose Acetate ◽  
Isoelectric Focusing ◽  
Boron Trifluoride ◽  
Serum Proteins ◽  
Ph Gradients ◽  
Specific Identification ◽  
Cellulose Acetate Membranes ◽  
Protein Staining

Abstract We applied a method previously described for treating cellulsoe acetate gel strips with boron trifluoride in methanol, to make them suitable for isoelectric focusing of proteins, to dry cellulose acetate membranes. The results obtained are shown to compare very favorably. Both wide and narrow pH gradients may be used and coupled either to conventional protein staining or to specific identification by immunofixation. Focusing ordinarily takes 90--120 min, is technically easy, and is economical in the use of ampholytes.

Fast Screening Method for Detection of Aflatoxin Contamination in Cottonseed Products

1972 ◽  
Vol 55 (5) ◽  
pp. 1114-1119 ◽  
Author(s):  
Alva F Cucullu ◽  
Walter A Pons ◽  
Leo A Goldblatt
Keyword(s):  
Detection Method ◽  
Screening Method ◽  
Aflatoxin Contamination ◽  
Contamination Level ◽  
Fluorescent Band ◽  
Total Analysis Time ◽  
Fast Screening ◽  
Total Analysis ◽  
Acidic Alumina ◽  
Benzene Extract

Abstract The detection method incorporates a 3 min blender extraction of the sample with aqueous acetonitrile, followed by partition of aflatoxins from an aliquot of the extract into benzene to effect a 4-fold concentration of the aflatoxins. A millicolumn (4 mm id × 20 cm long) filled with zones of acidic alumina (1.5 cm) and silica gel (9 cm) is placed in a portion of the benzene extract and the column is developed 5 min in chloroform-acetonitrile-2-propanol. Aflatoxins are detected by the presence of a sharp blue fluorescent band about 1 cm above the alumina zone when the developed column is examined under longwave ultraviolet light. Total analysis time is 15 min, and sensitivity of the method is in the range of 10–20 μg total aflatoxins (B1 + B2)/kg. An indication of the approximate contamination level can be obtained from the intensity of the fluorescent aflatoxin band.

Isoelectric focusing in agarose: classification of genetic variants of alpha 1-antitrypsin.

1983 ◽  
Vol 29 (2) ◽  
pp. 328-331 ◽  
Author(s):  
G J Buffone ◽  
B J Stennis ◽  
C M Schimbor
Keyword(s):  
High Resolution ◽  
Isoelectric Focusing ◽  
Genetic Variants ◽  
Virtual Absence ◽  
Agarose Gels ◽  
Limited Success ◽  
The Matrix ◽  
Alpha 1 Antitrypsin

Abstract Polyacrylamide has been the matrix of choice for isoelectric focusing owing to the virtual absence of electroendosmosis in this medium. Certain inherent limitations associated with polyacrylamide have prompted some investigators to use low-electroendosmosis agarose for isoelectric focusing, but with limited success thus far. We have developed a method for isoelectric focusing in agarose for the classification of alpha 1-antitrypsin variants. Sera are applied directly to agarose gels containing a pH 4-5 ampholyte mixture, focused for less than 1 h, and directly immunofixed. Resolution of major bands is equivalent to polyacrylamide, and Pi M subtypes can be distinguished without the use of a separator. This application demonstrates the high resolution of isoelectric focusing in agarose, a more practical and convenient matrix than polyacrylamide.

Analysis of the genetic variants of the human Gc system (VDBP) by isoelectric focusing in immobilized pH gradients

1982 ◽  
Vol 3 (6) ◽  
pp. 342-345 ◽  
Author(s):  
Hartwig Cleve ◽  
Walburgis Patutschnick ◽  
Wilhelm Postel ◽  
Johann Weser ◽  
Angelika Görg
Keyword(s):  
Isoelectric Focusing ◽  
Genetic Variants ◽  
Ph Gradients
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