Protein human serum fractionated

Figure 3. Fractionation of human serum proteins on Spherogel TSKSW 3000, The conditions were as in Figure 1. The analyses were made using (A) a 50-fiL injection loop with an analytical flow cell (B) a 100-L loop with a semipreparative flow cell or (C,D) a 500-L loop with a preparative flow cell.

There is no agreement on the exact magnitude of the ultrafiltrable or high-molecular-mass fraction of Ni(II) in human serum. Moreover, pronounced species variations in the proportions of ultrafiltrable and protein-bound serum nickel have been demonstrated [289],... 

El. Edelstein, C., Lim, C. T., and Scanu, A. M., On the subunit structure of the protein of human serum high density lipoprotein. I. A study of its major polypeptide component (Sephadex, fraction III). J. Biol. Chem. 247, 5842-5849 (1972). 

Most frequently, binding protein is added to the incubation mixtures as either serum or purified serum albumin. With human serum albumin, at equilibrium, the acceptor substrate will largely be protein-bound, when the bilirubin albumin molecular ratio is smaller than one (the dissociation constant of the first binding site of purified human serum albumin is approximately 7 X 10 M with 2 X 10 M for two additional binding sites) (J2). The first binding site of albumin, measured with rat serum, has a dissociation constant of about 5 X 10" M (M8). The unbound fraction will normally not exceed the very low solubility of the pigment. Addition of albumin to an alkaline solution of bilirubin, or its addition immediately after neutralization, prevents colloid formation, if the bilirubin albumin molecular ratio is smaller than one (B25). However, colloidal bilirubin, once formed, cannot be redissolved by the addition of albumin (B26). 

Subsequent work by Gardiner et al. showed that in a relatively complex mixture like human serum, the association between a metal and protein or ligand could be said to have been established when their Gaussian distribution coincide. This is more likely to be true if the elution volumes of the constituents are in the fractionation range rather than in the excluded volume. Examples of the usefulness of immunological techniques as an aid in identification of proteins are given. Necessary clean-up procedures are also suggested. 

Ldnnerdal, B., Stanislowski, A. G., Hurley, L. S. J. Inorg. Biochem. 12, 71 (1980) Woittierz, J. R. W. Elemental Analysis of Human Serum and Human Serum Protein Fractions by Thermal Neutron Activation, Netherlands Energy Research Foundation Report, ECN 147, January 1984... 

Figure 6.10 Binding of toxicants to blood proteins (a) Double-reciprocal plot of binding of rat serum lipoprotein fraction with four insecticides. Insert illustrates magnitude of differences in slope with Scatchard plot, (b) Scatchard plot of binding of salicylate to human serum proteins. (Sources (a) Skalsky and Guthrie, Pest. Biochem. Physiol. 7 289, 1977 (b) Moran and Walker, Biochem. Pharmacol. 17 153, 1968.)...

A rapid two-dimensional separation of a microspot of protein solution under high field intensity is a new application of the two-dimensional principle. The various fractions emerge as a series of rays out of the application zone and hence the designation star electrophoresis which was introduced by us in connection with work on human serum proteins (P4). A similar discontinuous procedure was already mentioned by Strain in 1951 (S4, S5) for the separation of metal ions and recently used by Pucar (P8) and applied by Keler et al. (K1) to human serum. 

Different physical techniques can also be used for prefractionation of samples to be analyzed by 2-D gels. Corthals et al. have fractionated human serum by an electrokinetic technique [67], Fountoulakis et al. have used chromatofocus-ing as a prefractionation step in the analysis of low-abundance proteins in H. 

In a follow-up study, the same authors examined the applicability of the same device for relevant protein samples and investigated the main contributions to band broadening [82]. As a consequence of the small depth of the beds, zone spreading caused by Joule heating was shown to be negligible (see Sect. 3.1.1). Cross fields of up to 100 V/cm were applied for the separation of human serum albumin, ribonuclease A and bradykinin. The feasibility of fraction collection was demonstrated with four collected fractions of a whole rat plasma sample. Off-line analysis of these four isolated fractions by CE indicated the separation of serum albumins and globulins. 

The iron-binding protein of serum transferrin was found in fraction IV-3,4 of human plasma when the plasma was fractionated by low temperature ethanol fractionation procedures (31, 116). By further subfractionations, serum transferrin could be concentrated in Cohn fraction IV-7 (30, 125, 126). Cohn (30) first reported the properties of the isolated protein, which he called the 3i metal-binding protein since the protein had been found to bind copper, and possibly zinc, as well as iron. Holm-berg and Laurell (66) proposed that the protein be called transferrin on the basis that the principal function of the protein was associated with the transport of iron in serum and that it was not the major copperbinding protein in human serum. 

Amino acid composition of the serum proteins. II. Fractionation of human serum proteins by cellulose ion-exchange chromatography and comparative amino acid composition of the fractions. Ibid., 19, 451 (1958). With S. Keller. 

Plasma Protein Binding (PPB). The fraction bound to plasma proteins (%PPB) can be easily calculated from binding affinity to HSA under the assumption that binding occurs mainly to the Human Serum Albumin (HSA). Thus we can ignore differentiation of both these properties. A dataset of 94... 

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