Simplified Determination of Fibrinogen Subfractions by Glycine

1979 ◽  
Author(s):  
S. Sasaki ◽  
K. Kito
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Human Plasma ◽  
Gel Electrophoresis ◽  
Biological Properties ◽  
Strength Analysis ◽  
Simple Method ◽  
Molecular Fraction ◽  
The Difference

Although it has been suggested by some investigators that human plasma fibrinogen is composed of two or more subfractions, no simple method of determining these fractions individually has been available for clinical use. We found that precipitation by glycine at certain ionic strength was suitable for this purpose. One volume of human plasma was added to 10 volume of 2.3 M glycine solution of varying ionic strength. Analysis of the precipitates obtained at various Ionic strength by means of SDS-gel electrophoresis gave the following results: The precipitate obtained at ionic strength 0.2 gave single band. Its molecular weight was 360,000, clottability 85%. The supernatant was devoid of this fraction. The precipitate obtained at ionic strength 1.4 gave two banda corresponding to molecular weight of 360,000 and 325,00O respectively. The clottability of the precipitate was 85%. The supernatant contained no significant amount of clottable protein. It is therefore possible to determine the concentrations of high molecular fraction of fibrinogen and of total fibrinogen in plasma using the precipitate at ionic strength 0.2 and the precipitate at ionic strength 1.4 respectively. The difference of the two values gives the concentration of low molecular fraction of fibrinogen. Biological properties of the two fractions were also studied.

Preparation of Human Plasminogen Suitable as Substrate for Quantitative Determination of Plasminogen Activators

1966 ◽  
Vol 15 (03/04) ◽  
pp. 511-518
Author(s):  
W Berg ◽  
K Korsan-Bengtsen ◽  
J Ygge
Keyword(s):  
Ionic Strength ◽  
Quantitative Determination ◽  
Spontaneous Activity ◽  
Human Plasma ◽  
Gel Filtration ◽  
Plasminogen Activators ◽  
Simple Method ◽  
Neutral Ph ◽  
Human Plasminogen

SummaryA simple method for preparation of plasminogen with low spontaneous activity and soluble at a neutral pH and at physiological ionic strength is described. Euglobulin made from fresh, oxalated, BaSO4-adsorbed, human plasma was first purified by means of gel filtration on Sephadex G-200. After gel filtration, further purification and concentration was done on DEAE-sephadex A-50. The activity was 100-130 casein units per mg tyrosine.

Partial purification of a boar sperm membrane protein inducing sperm agglutinating antibody

1990 ◽  
Vol 10 (2) ◽  
pp. 131-139
Author(s):  
Oyewole Adeyemo ◽  
E. O. Okegbile ◽  
O. O. Olorunsogo
Keyword(s):  
Molecular Weight ◽  
Membrane Protein ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Partial Purification ◽  
Boar Sperm ◽  
Sperm Membrane ◽  
Triton X

For the development of immunological contraception, attention is being concentrated on the possibility of using a sperm membrane antigen. Boar sperm membrane was extracted with triton-X 100 and fractionated by Sephadex G-150 column chromatography. The glycosylated and nonglycosylated portions of protein peaks from the gel filtration were obtained by fractionating on concanavalin A-Sepharose and eluting the bound protein with 0.3 M methyl mannoside. A glycosylated fraction was found to induce sperm agglutinating antibodies in rabbit. The partially purified protein has a molecular weight of 30 kilodaltons, as determined by sodium dodecyl polyaccyrlamide gel electrophoresis. Further work is planned on the histochemical determination of the origin of this protein and species cross-activity of the antibody.

scholarly journals SIMPLE METHOD FOR OPTIMIZATION OF HEPARIN AND ENOXAPARIN DETECTION USING AGAROSE GEL ELECTROPHORESIS

2019 ◽  
pp. 12-15
Author(s):  
Ahmad Abdulrahman Almeman
Keyword(s):  
Molecular Weight ◽  
Toluidine Blue ◽  
Gel Electrophoresis ◽  
Optimization Procedure ◽  
Solvent Mixture ◽  
Acid Mixture ◽  
Low Molecular Weight ◽  
Simple Method ◽  
Electrophoresis Method ◽  
Tools And Techniques

Objective: A simple gel electrophoresis method for low-molecular-weight heparins (LMWH) is required for use in a variety of laboratories to allow further identification and purification. This study aimed to optimize the detection of heparin and enoxaparin (low-molecular-weight heparin by gel electrophoresis. Methods: Several gel electrophoresis conditions were tested to optimize the detection of enoxaparin by using a simple method with a modified Volpi’s approach. Multiple gel thicknesses, voltage settings, and enoxaparin concentrations were tested in the optimization procedure. Enoxaparin was purchased from a local supplier as pre-filled pharmaceutical injections. Highly purified 0.5% and 1.0% agarose gels were prepared and a series of enoxaparin concentrations was added to both gels for comparison and optimization. The 0.2% toluidine blue stain was prepared by the addition of 1 ml in an ethanol-water-acetic acid mixture (50:49:1; v/v/w). The staining process comprised two steps: first, toluidine blue was added for 30 min and destained overnight in the solvent mixture. Subsequently, the following morning, the second step was conducted, in which the gel was restained for 30 min with the same concentration of toluidine blue. We continued to stain the gel until the bands were visible. Results: The gel electrophoresis results showed that clearest and sharpest bands were obtained using 65–75 mAh and 85 V settings. At 95 mAh, the bands were slightly washed out. Conclusion: This study successfully facilitated the detection of enoxaparin, a LMWH, and heparin in the laboratory by using simple tools and techniques available in most laboratories.

RADIOIMMUNOASSAY OF OESTRONE AND OESTRADIOL IN HUMAN PLASMA

1972 ◽  
Vol 69 (3) ◽  
pp. 567-582 ◽  
Author(s):  
Yvonne Emment ◽  
William P. Collins ◽  
Ian F. Sommerville
Keyword(s):  
Menstrual Cycle ◽  
Human Plasma ◽  
Simple Method ◽  
Healthy Women ◽  
Theoretical Error ◽  
Dextran Coated Charcoal ◽  
Coefficients Of Variation ◽  
The Mean

ABSTRACT A simple method for the determination of oestrone and oestradiol in human plasma is described and evaluated in terms of theoretical and practical errors. The oestrogens are separated on columns of Sephadex LH 20 and, after equilibration with antiserum, the unbound steroid is removed with dextran-coated charcoal. The mean total random theoretical error is calculated to be 22 %; the coefficients of variation of replicate analyses on pooled female plasma were from 13–18 % for both steroids; in male plasma the values were 17 % for oestradiol and 27 % for oestrone. The concentrations of oestradiol, expressed in pg/ml ± sd) in samples collected from healthy women during the menstrual cycle were 69 ± 56 (days 1–10); 126 ± 66 (days 11–20) and 99 ± 54 (days 21–30). Corresponding values for oestrone were 119 ± 46, 156 ± 41 and 156 ± 27.

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