Protein bovine fibrinogen

Protein samples used to conduct a protein transfer were bovine serum albumin (BSA, 67,000 daltons), chicken egg ovalbumin (Ovalb, 45,000 daltons), and human serum cryo-precipitate. These agents were obtained from plasma containing a mixture of proteins including Fibrinogen (340,000 daltons), human serum albumin (HSA, 67,000 daltons), and immunoglobulin G (IgG, 47,000-56,000 daltons). The cryo-precipitate was diluted with 20 ml of buffer solution prior to separation. 

They experimented with four other proteins as carriers rabbit serum albumin, bovine serum albumin, bovine fibrinogen fraction I, and bovine )8-globulin fraction III. The structurally related derivatives of DDT and malathion, DDA, and 0,0-dimethyl S-carboxy-carboxyethyl phosphoro-dithioate (malathion half ester), respectively, were used as the specific haptens attached to these carrier proteins. These compounds contain free carboxyl groups, which when they reacted with thionylchloride, provide a means of coupling of the hapten to the amino groups of the protein carrier. 

The spectrophotometric evidence reviewed above for the binding of a proportion of the phenolic hydroxyl groups of the tyrosine residues of native proteins is supported by work on the action of tyrosinase on proteins. Sizer (1946) found that this enzyme oxidizes the tyrosine residues in native trypsin, pepsin, chymotrypsin, casein, peptone, insulin, and hemoglobin. Native ovalbumin, human and bovine serum albumin, tobacco mosaic virus (nucleoprotein), human y- and bovine /3-globulins, and bovine fibrinogen are not susceptible to tyrosinase, but become so after tryptic digestion. It was shown (Sizer, 1947) that for the proteins which are oxidized by tyrosinase in the native state, the observed reaction does indeed occur with the intact proteins and does not require preliminary degradation to tyrosine peptides or free tyrosine. The kinetics of the oxidation of tyrosine by tyrosinase have been studied spectropho-tometrically (Mason, 1948 etc.). 

Fia. 68. The electrophoretic mobility of bovine fibrinogen and fibrin plotted against pH 10 mg per milliliter protein 0.1 ionic strength 20% urea (10% urea below pH 5.9). Open circles fibrinogen. Solid circles fibrin (Mihalyi, 1950b). 

The effects of conditioning layers of two important blood serum proteins, albumin and fibrinogen were investigated. Protein adsorption was studied using bovine serum albumin (BSA) and fibrinogen (F) from Sigma. The samples were incubated for 3 h at 37°C in solutions of albumin (1 mg/mL) and fibrinogen (0.2 mg/mL) prepared in phosphate buffered saline (PBS, 0.01 M phosphate buffer, 0.0027 M KC1, 0.137 MNaCl, pH 7.4). After the incubation period, the samples were rinsed 3 times with PBS and analyzed by the various surface characterization techniques. 

Table 1. Atomic percentages for the 5 W PEO-like and Ag/PEO-like films, following incubation in the protein solutions, bovine serum albumin and fibrinogen. In this figure BSA represents bovine serum albumin and F represents fibrinogen.18...

Proteins used in this study are albumin (bovine, crystalline, Nutri-tritional Biochemical Co.), y-globulin (bovine. Fraction II, Nutritional Biochemical Co.), prothrombin (bovine. Fraction III-2, Nutritional Biochemical Co.), I-y-globulin and I-fibrinogen (New England Nuclear Co.), and I-albumin (Squibb Co.). 

No definitive study of the effects of protein carrier on response is available, and many carrier proteins have been used, including globulins, albumin, hemocyanin, thyroglobulin, and fibrinogen. The optimal number of haptens, or the epitope density, is also controversial, but a density of 8 to 25 haptens per bovine serum albumin molecule is probably optimal (12),... 

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