Albumin, electrophoretic analysis

Figure 7.7. Association isotherms of micelles (TX 100) or the protein bovine serum albumin on azobenzene-modified poly (sodium acrylate) in dilute water solution, as measured from capillary electrophoretic analysis. Closed symbols dark-adapted samples, open symbols samples exposed to UV light (365 nm). (a) Polymer, as shown in Fig. 7.1, with n = 5 and x=7%, (b) n = 12 and x=3%.

Two-dimensional electrophoretic analysis of tear proteins followed by immunoblotting has shown that the major components are lactoferrin, lysozyme, albumin, secretory IgA, and five tear-specific proteins. There are significant differences in the amounts of lactoferrin and two kinds of tear-specific proteins differ between the sexes. A comparison of the two-dimensional electrophoretic patterns of tear proteins from patients with conjunctivitis with normal individuals shows reductions in the levels of IgA, lactoferrin, and tear-specific proteins whilst the intensity of albumin is increased in addition, haptoglobin and IgG are present in the patient samples. 

Detection was with Coomassie Blue. Lanes A and B, sample containing 2.95 mg protein/ml and 70 U/ml of a-amylase estimated by 3,5-dinitrosalicylic acid [8]. Lane C, 1 myosin (205,000), 2 3 galactosidase (116,000), 3 phosphorylase (97,000), 4 bovine serum albumine (66,000), 5 egg serum albumine (45,000), 6 carbonic anhydrase (29,000). The author thanks Dr. M. JiresovA for performing the electrophoretic analysis of a-amylase. 

This factor together with a dependence of precipitation on the concentration of electrolyte precludes ready quantitative separation of components of different isoelectric point in protein mixtures such as serum (118, 124), Electrophoretic analysis has revealed that one precipitation with SDS achieves only partial separation of the components of a synthetic mixture of purified albumin and globulin buffered in the interisoelectric region (118). 

Fig. 3. Electrophoretic analysis of serum albumin—sodium dodecyl sulfate mixtures at 1°C. Figures between patterns represent experiment numbers. In all experiments 0.3% detergent was present except in experiment 1 which contained only pure protein. In experiments 2 to 6 the protein concentration decreased as follows 2.0%, 1.5%, 1.0%, 0.8% and 0.6%. Since all experiments were run for the same time (14,400 sec.) at essentially the same field strength (4.7 v./om.), constancy of mobilities of components labeled A, 1,2, and D may be observed by comparing directly displacements of respective boundaries from the common origin (starting boundary characterized by the pair of vertical lines). Arrows denote direction of migration right, descending left, ascending boundary. From Putnam and Neurath (117).

In support of this hypothesis, Putnam and Neurath (117) were able to demonstrate that the binding of tingle detergent ions with individual protein groups took place stoichiometrically. Electrophoretic analysis indicated that the two serum albumin-SDS complexes at pH 6.8 had the same constant composition as the complexes that precipitate at pH 4.5. Similar calculations (81, 117) indicated that the number of moles of alkylbenzene sulfonate strongly bound in neutral solution by egg... 

Electrophoretic analysis of this serum gave the following results, in terms of per cent total refractive area albumin 32.9%, o-globulins 6.2%, /1-globulins 11.8%, and T-globuIins 50.2 %. 

Seibert et al. (317) recently reported results of electrophoretic analysis of the serum proteins in 23 patients with carcinoma originating from a variety of sites. The albumin was lowered, airglobulins were usually increased, the 7-globulins were usually low, although in some instances, particularly those with jaundice due to metastatic involvement of the liver, elevated 7-globulins were the rule. Our own experience indicates that secondary infections play an important part in affecting plasma protein patterns in carcinoma patients. In those cases associated with... 

Analysis of in vivo-formed pellicle by a combination of electrophoretic separation and MALDI-TOF showed the presence of intact histatin 1, cystatin SN, statherin, lysozyme, albumin and amylase [15, 39], In addition, intact cytokeratins 13 and 15 and calgranulin B were identified as components of the salivary pellicle layer for the first time using MALDI-TOF mass spectrometry [15]. Calgranulin B has been shown to be a component of saliva and gingival crevicular fluid [15]. The identification of cytokeratins in the salivary pellicle layer points to the oral epithelium as one of the sources of proteins adsorbed on the tooth surface. 

Lipids circulate in blood as constituents of the lipoproteins or bound to albumin (free fatty acids). Electrophoretic separation of plasma proteins on paper is a relatively easy and convenient method for the semi-quantitative analysis of lipoproteins. Staining the strips with dyes which only respond to lipids reveals three well-defined bands when normal plasma or serum is analyzed. The a-lipoprotein migrates the farthest from the origin. The pre-j8, previously called the a-2-hpoprotein, and the jS-Hpoprotein, are closer to the origin. In the post-prandial normal serum, and in some hyperlipemic sera, after fasting, a fourth band can be seen at the origin. This band is due to neutral fat particles with a small protein content (i.e. chylomicron fraction). 

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