Serum albumin, bovine preparation

Prepare competitor protein buffer detergent solution 4 mg/mL normal goat globulin or other competitor protein, (such as fetal calf serum, bovine serum albumin, bovine plasma, and so forth), and 0.1% saponin (Sigma, St. Louis, MO) in phosphate-buffered saline (NGG-sap-PBS). 

In each of these methods, undiluted serum, urine, acid-deproteinized milk, or a buffered saline extract of muscle was mixed with sulfamethazine-horseradish peroxidase and added to antibody-coated wells of a microtiter plate. A sulfamethazine-bovine serum albumin conjugate prepared by the glutaraldehyde procedure (56) was used for antibody production. Results showed that screening of serum was of value since sulfamethazine concentrations in serum directly correlated with those in swine tissues. Thus, for example, a level of 100 ppb of sulfamethazine in... 

Our results indicate the importance of using a unified protein standard for calibration and traceability. Having no RM to make a comparison against, we used bovine serum albumin (Fluka, cat. No. 05470) which was traceable to the NIST SRM 927c (Total Protein Standard) bovine serum albumin. Both preparations can be used for traceability. 

FIGURE 7.10 Dependence of the resolution on the sample volume. A preparative Superformance column 1000-200 (bed volume 20 liters) packed with Fractogel END BioSEC (S) (bed height 63 cm) was loaded with 60 ml (top) and 300 ml of a mixture of bovine serum albumin (5 mg/ml), ovalbumin (5 mg/ml), and cytochrome c (3 mg/ml) (bottom) (20 m/VI sodium phosphate buffer, 0.3 M NaCI, pH 7.2 flow rate 100 ml/min corresponding to 19 cm/hr). When the sample volume is 300 ml the separation efficiency for BSA and ovalbumin is similar. Thus the column can be loaded with larger sample volumes, resulting in reasonable separations. 

Protein concentration can be determined using a method introduced by Bradford,4 which utilises Pierce reagent 23200 (Piece Chemical Company, Rockford, IL, USA) in combination with an acidic Coomassie Brilliant Blue G-250 solution to absorb at 595 nm when the reagent binds to the protein. A 20 mg/1 bovine serum albumin (Piece Chemical Company, Rockford, IL, USA) solution will be used to prepare a standard calibration curve for determination of protein concentration. The sample for analysis of SCP is initially homogenised or vibrated in a sonic system to break down the cell walls. 

Microspheres were prepared from carboxymethyl chitosan and alginate by emulsion phase separation. The encapsulated bovine serum albumin was... 

Miyake et al reported an ELISA method for the determination of pesticide residues in the aquatic environment. The polyclonal antibody and three monoclonal antibodies of acifluorfen were prepared by immunization of rabbits and mice with acifluorfen-bovine serum albumin conjugates. The polyclonal antibody reacted with acifluorfen at concentrations of 1.5-800 mg L , while the monoclonal antibodies reacted with acifluorfen at concentrations of 1.5-144 mg L . Among three monoclonal antibodies, AF 75-144 reacted with chlornitrofen, which did not react with the other two antibodies. It seems that the ELISA method is effective for the determination of herbicide residues in the aquatic environment. 

We used an anti-DNA antibody as an exploratory model system. The antibody was monoelonal from mouse sourees and its subelass was IgM. Mouse IgG (MW 1.5 x 10 Da) and IgM (MW 9 X 10 Da) antibodies from normal plasma, and bovine serum albumin were used for the eontrol measurements. To prevent the nonspeeilie adsorption of proteins to the uneovered, bare Au site in the modified eleetrode surfaee, the DNA-modified eleetrode prepared by the standard proeedure was further treated with aqueous 2-mercaptoethanol solution and was used for the measurements. 

Membranes offer a format for interaction of an analyte with a stationary phase alternative to the familiar column. For certain kinds of separations, particularly preparative separations involving strong adsorption, the membrane format is extremely useful. A 5 x 4 mm hollow-fiber membrane layered with the protein bovine serum albumin was used for the chiral separation of the amino acid tryptophan, with a separation factor of up to 6.6.62 Diethey-laminoethyl-derivatized membrane disks were used for high-speed ion exchange separations of oligonucleotides.63 Sulfonated membranes were used for peptide separations, and reversed-phase separations of peptides, steroids, and aromatic hydrocarbons were accomplished on C18-derivatized membranes. 

Materials. Microspherical PGG glucan (Adjuvax, Alpha-Beta Technology, Worcester, MA) was prepared from Saccharomyces cereviseae strain R4 cells (11). Zymosan, cytochrome c (cyt c), bovine serum albumin (BSA), yeast alcohol dehydrogenase (ADH), Complete Freunds Adjuvant (CFA) and Incomplete Freunds Adjuvant (IFA) were purchased from Sigma Chemical Co. (St. Louis, MO). 

Preparations of PEG-modified proteins. A. SC-PEG (1 g, 0.2 mmol) was added to a stirred solution of Bovine Serum Albumin (BSA) (100 mg, 1.5 x 10 6 mol) in 0.1 M sodium phosphate, pH 7.8 (60 mL). Sodium hydroxide (0.5 N) was used to maintain pH 7.8 for 30 min. The excess of free PEG was removed by diafiltration using 50 mM phosphate buffered saline. Approximately 30 amino groups of the native protein were modified as determined by trinitrobenzenesulfonate (TNBS) assay (28). The same degree of modification was obtained when the experiment was repeated under identical conditions using SS-PEG instead of SC-PEG. 

Fig. 3.9 SEM images of aerogels. They were prepared from initial gels synthesized by adding 10wt.% THEOS to an aqueous solution containing 1 wt.% bovine serum albumin at the various pHs shown underthe pictures. (Unpublished results obtained with the participation of N. Shipunova and D. Fomin).

We have observed that such proteins as CaM and bovine serum albumin (BSA) can be developed at the air-water interface to form monolayer protein films. In previous works, the developed BSA monolayer was stabilized by cross-linking with a bifunctional reagent immediately after the preparation of protein monolayer. The BSA thin film thus prepared can be employed as a passive material, e.g., an ultrathin protein film for a matrix of enzyme-linked immunosorvent assays. 

In this method the keyhole limpet haemoglobin conjugate was prepared as follows Keyhole limpet haemocyanin (KLH, Calbiochem, La Jolia, CA) and bovine serum albumin (BSA, BDH Chemicals) were coupled to the adduct (2), derived from 6-bromohexanoic acid and monoquat (3), via a carbodiimide reaction, as reported previously by Niewola et al. [184], The resulting conjugates contained 662mol of Paraquat per mole of KLH and 15mol of Paraquat per mole of 6-bromohexanoic acid. The amount of Paraquat bound to the protein was determined by spectrophotometric dithionite assay for Paraquat and the protein concentration was established by a standard Lowry test. 

Antibody Production A thick emulsion of the immunogen (clonazepam-bovine-serum-albumin-con-jugate) is prepared employing complete Freund s adjuvant and two New Zealand white female rabbits are immunized intradermally at multiple sites with the immunogen emulsion. The animals are then administered... 

The resulting product (II) is subsequently coupled to bovine-serum-albumin in a glycerol-w ater mixture in the presence of dicyclohexylcarbodiimide. The mixture is incubated overnight at 4°C, and the protein-hapten complex is dialysed against distilled water thereby causing its purification. Conjugation of the respective barbiturate to the protein carrier, comparison of the barbiturate BGG-conjugate to control BGG-solution and preparation of 14C-pentobarbital sodium are carried out respectively. 

Preparation of Antiserum The barbiturate-bovine-serum-albumin conjugate is duly emulsified with an equal volume of complete Freund s adjuvant and New Zealand albino rabbits are subsequently im-... 

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