Whole serum

The immunoglobulin fraction from each bleed was purified by use of a recirculating isoelectric focusing (RIEF) technique (Bier et al. 1979). The whole serum was diluted 1 3 with urea, to yield a final urea concentration of 3 M, and then desalted by electrodialysis. The urea was added to prevent precipitation under hypotonic conditions. Ampholine (1 percent w/v, pH 3.5 to 10, LKB... 

The affinity and cross-reactivity of the whole serum and Fab fragments were determined using equilibrium dialysis for the affinity determination and RIA for the cross-reactivity studies. The average intrinsic affinity constant (Ko) of the antibody (Nisonoff and Pressman 1958) changed very little throughout the... 

Purified antibodies are usually prepared from whole serum by affinity chromatography. Concentration of affinity-purified specific antibodies is normally specified by the manufacturer in the antibody data sheet. When starting with a new... 

B1 ood Whole serum or bl ood Acidification solvent GC/ECD extraction clean-up with concentrated sulfuric sulfuric acid No data 84.7 (serum) 85.1 (whole blood) Waliszewski and Szymczynski 1991 ... 

Since the early delipidation procedure was applied initially to the whole serum (815) and later to the isolated serum lipoproteins (818), many other methods of delipidation have been reported [see reference (816) for review], employing mixtures of organic solvents (ethanol-ethyl ether chloroform-methanol, acetone, etc.) or detergents (sodium dodecyl or decyl sulfate, Triton X-100, Nonidet, etc.). Techniques for delipidation have not been standardized, nor is there a comprehensive comparative assessment of the various proposed methods presently avail-... 

Denison, M.S., Rogers, W.J., Fair, M., Ziccardi, M., Clark, G., Murk, A.J., Brouwer, A. (1996). Application of the CALUX bioassay system for the detection of dioxin-like chemicals (Ah receptor ligands) in whole serum samples and in extracts fiom commercial and consumer products. Organohalogen Compounds 27 280-284. 

We now may examine the relevance of surface tension measurements to interactions of cells in contact with serum or plasma. The surface tensions reported in Table I for the unfiltered liquids are much lower than those of the ultrafiltrates, at least in part, because of the adsorption of proteins having molecular weight > 20,000 at the liquid-air interface. And the measurements made on the ultrafiltrates are likely to be a good approximation to the zero-time surface tensions of the whole serum and plasma. The solutes that remain in the solution after ultrafiltration evidently are of a relatively low level of surface activity, and do not affect the surface tension to any greater extent than would be expected from their volume fractions. 

Monoclonal antibody therapy (MAT) makes use of all the major features of the immune response. It involves vaccination/ immunization, albeit in experimental animals, to induce the desired specific immune response. It exploits the high specificity, selectivity, and affinity of the antibody CDR toward the target antigen to be recognized, highlighted, inactivated, or eliminated, using the characteristics of the Fc portion of an immunoglobulin to facilitate the means for such inactivation or elimination and for selection of appropriate effector mechanisms. Finally, MAT represents a modern form of serotherapy, in which parenteral administration of whole serum or Ig preparations has been replaced by recombinant antibody molecules of a defined specificity. 

Sephadex treatment. A sterile column (2.5 x 100 cm) containing about 50 g dry weight Sephadex G-50 (coarse) is well washed with 5 1 sterile 0.85% saline. The Sephadex slurry may be sterilised by autoclaving for 15 min at 121 °C before packing. 200 ml of whole serum may be applied to such a column and eluted with saline at 4-5 ml/min. The excluded fraction elutes between 200 and 400 ml. 

Constituent Whole serum Sephadexed serum Dialysed serum... 

Fig. 6.2. Prominent parasite antigens and host components in Echinococcus granulosus sheep cyst fluid (SCF). Labelled fluid was immunoprecipitated with lane 1, normal rabbit serum lane 2, rabbit anti-SCF lane 3, preabsorbed rabbit anti-SCF lane 4, rabbit anti-sheep whole serum. Antigens were reduced prior to electrophoresis. The major parasite antigens are shown arrowed at Mr 38000, 20000, 16000 and 12000. D.F., diffusion front. (Reprinted with permission from Molecular and Biochemical Parasitology, 25, Shepherd, J. C. McManus, D. P., Specific and cross-reactive antigens of Echinococcus granulosus hydatid cyst fluid, 1987, Pergamon Journals Ltd.)...

For polyclonal antibodies, negative reagent controls should be a dilution of immunoglobulin fractions or whole serum of normal/ non-immune serum of the same animal source. Again, the negative reagent control should be applied in the same concentration as the test antibody, and the same diluent should be used. 

Y. Kita, Y. Miura, J.-i. Furukawa, M. Nakano, Y. Shinohara, M. Ohno, A. Takimoto, and S.-I. Nishimura, Quantitative glycomics of human whole serum glycoproteins based on the standardized protocol for liberating IV-glycans, Mol. Cell. Proteomics, 6 (2007) 1437-1445. 

Preparation of Immunoglobulin Fractions from Whole Serum... 

Antibody—either diluted whole serum or IgG fraction (0.1 ml)... 

Antiserum is that part of the serum containing antibodies. It may be prepared from whole serum by a series of ammonium sulfate precipitation steps, where successively lower concentrations of ammonium sulfate generate a precipitate that is centrifuged and resuspended in buffer.4 The antiserum is then dialized or affinity-purified, and assayed for total protein. 

The immunogen dissolved in saline or distilled water (sterile) should not exceed a maximum concentration of about 75 mg/ml to obtain stable emulsions whole serum should be diluted at least 50 times. The failure to make a proper emulsion leads to disappointing results of immunizations. A simple way to prepare stable emulsions (Herbert, 1973) is to place the two solutions (same volumes)... 

In the above methods, the separated albumin (or whole serum if total protein is to be determined) is reacted with excess copper in approximately 1 N alkali. A macromethod is also available which utilizes the capacity of albumin to react with copper(II) in a mole ratio of 1 1 (K32). This is an amperometric titration of blood serum (1 ml) in 0.1 M ammoniacal am-moniiun nitrate pH 9.2 with CuS04 (4.8 X 10 M). Under these conditions human y-globulin did not react, so that the fair agreement by this method with results obtained by a biuret method after sulfite fractionation (K32) is probably the result of a balance of errors. In view of the interest in the small fraction of albumin-boimd copper in hepatolenticular degeneration, and the paucity of information on the unique binding reaction presumably with N-terminal aspartyl residues, further investigation would be valuable. 

Fluorescent band with emission maximum at 425 nm, distinct from albumin and NADH Fluorescence in whole serum before and after addition of CK assay mixture... 

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