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Topic:Electrophoresis
Electrophoresis is a laboratory technique used to separate and analyze macromolecules, such as proteins and nucleic acids, based on their size and charge. In equine research, electrophoresis is often applied to assess protein profiles in horse serum or plasma, aiding in the diagnosis and monitoring of various health conditions. This method allows for the identification of specific protein patterns associated with diseases, nutritional status, and physiological changes in horses. Electrophoresis can be used to detect abnormal protein levels and to evaluate the presence of specific proteins that may indicate underlying health issues. This page gathers peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings related to electrophoresis in equine health diagnostics and research.
Origin an importance of increased alkaline phosphatase activity in peritoneal fluids of horses with colic. The origin of increased alkaline phosphatase (ALP) activity in peritoneal fluid (PF) of horses with clinical signs of abdominal pain was investigated to determine the usefulness of measuring ALP in PF in the diagnosis of small intestinal injury. The ALP isoenzymes in PF from 10 clinically normal horses and from 50 horses with clinical signs of acute abdominal pain were analyzed for their sensitivities to inhibition by L-phenylalanine, L-homoarginine, and levamisole and to inactivation by heat (56 C, 15 minutes). The enzymes also were discriminated by their patterns of migration during polyacry...
Fibrinolytic activity without fibrinogenolysis during long-distance racing in horses. Fourteen horses were studied during a 157-km endurance ride. Two humans who ran the 157 km were also evaluated at the finish. Fibrin monomer samples were examined by two-dimensional gel electrophoresis and sodium dodecyl sulfate polyacrylamide gel electrophoresis. Two major species of horse Beta-chain with higher molecular weights and different isoelectric mobilities than human beta-chain were observed. Horse alpha-chains had higher molecular weights than human alpha-chains but similar alpha-chain heterogeneities. Mean euglobulin lysis time (ELT) in the horses was accelerated to similar levels...
Equine marker genes: Polymorphism for soluble erythrocyte malic enzyme. Polymorphism of equine erythrocyte malic enzyme is detactable on starch gel electrophoresis. The frequency of ME1S was 0.06 in 667 Standardbred and 0.09 in 85 Thoroughbred horses. No genetically determined electrophoretic variation in soluble malate dehydrogenase was detected.
Antigen-antibody crossed electrophoretic studies and quantitative comparisons of serum transferrin types in horses. Selected transferrin phenotypes from 14 horses were investigated by antigen-antibody crossed electrophoresis. Horse sera were subjected to starch gel electrophoresis followed by right angle electrophoresis in agarose gels containing rabbit produced anti-horse transferrin. This technique gave an additional zone in the front as compared with 2 transferrin zones seen after ordinary starch gel electrophoresis. Comparisons of transferrin concentrations in horse sera were performed by an immunodiffusion technique. Values were related to a chosen reference serum. A total of 372 horses (210 Norwegian ...
Fractionation and partial characterization of alpha-1-protease isoinhibitors of horse. The principal alpha-1-protease inhibitor of horse was fractionated by classical methods and analysed with a modified fibrinogen-agarose gel electrophoretic method of high sensitivity and resolving power. Starting with an electrophoretically homogeneous inhibitor in unfractionated serum, two isoinhibitor bands became apparent after fractionation with (NH4)2SO4 and DEAE-cellulose DE-52 ion-exchange chromatography. The isoinhibitors differed in electrophoretic migration and in the elution pattern from Sephadex G-100 gel filtration, but possessed identical antigenic determinants and enzyme specifi...
Characterization of equine alkaline phosphatase isoenzymes based on their electrophoretic mobility by polyacrylamide gel disc electrophoresis. Alkaline phosphatase isoenzymes of equine tissues, peritoneal fluid, and serum were characterized by their electrophoretic mobilities, using polyacrylamide gel disc electrophoresis. The alkaline phosphatase isoenzymes in liver, kidney, spleen, small intestine, placenta, bone, small colon, and large colon tissue samples were extracted and separated by electrophoresis. The resulting isoenzyme mobilities and spectrophotometric scans were evaluated for their tissue specificity and for their possible use in determining the tissue contribution of alkaline phosphatase to serum and peritoneal fluid. T...
Antibody activities of immunoglobulins in anti-leptospiral horse sera. Antileptospiral sera from hyperimmunized horses were fractionated by gel filtration on Sephadex G-200 or by starch block electrophoresis. The fractions were examined quantitatively for leptospiricidal, agglutinating and complement fixing activities. The leptospiricidal activity was higher in the 78 globulin fraction than in the 19S globulin fraction, while the agglutinating activity was shared by both the fractions being higher in the 19S fraction. Complement fixing activity was found evenly in both the fractions. Leptospiricidal and complement fixing activities were higher in gamma-globulin t...
Assembly of intra- and interspecies hybrid apoferritins. An intraspecies hybrid apoferritin was assembled by mixing subunits of horse heart ferritin, which consists mainly of H-type subunits, and horse spleen ferritin, in which L-type subunits predominate. Interspecies hybrid apoferritins were reconstituted from subunits of human liver-horse spleen ferritins and from rat liver-horse spleen ferritins. All the hybrid ferritins migrated as single zones with electrophoretic mobilities intermediate between those of the parent ferritins. Isoelectric focusing data and immunological patterns were consistent with the view that the reassembled apoferritins we...
Variation of acidic prealbumins in the donkey (Equus asinus). Starch gel electrophoresis of 55 donkey serum samples revealed three prealbumin (Pr) phenotypes temporarily designated Pr M, Pr MT and Pr T. The distribution was in agreement with a genetic theory of two codominant alleles of frequencies, PrM = 0.87 and PrT - 0.13. Variation was also observed for proteins migrating with the same rate as the Xh zones in the horse.
Equine marker genes. Polymorphism for transferrin alleles, TfF1 and TfF2, in Thoroughbreds. The equine transferrin F variant is distinguishable into two types, F1 and F2, on alkaline polyacrylamide gel electrophoresis. Gene frequencies in 63 related Thoroughbreds are 0.39 and 0.19 for TfF1 and TfF2, respectively. In contrast the frequencies for these two alleles in 375 related Standardbreds is 0.00 and 0.59.
Protease inhibitor system in horses: classification and detection of a new allele. A method of horizontal thin layer polyacrylamide gel electrophoresis at acid pH has been developed for the separation of the prealbumins in equine plasma. Using this method, it has been possible to split the S allele into two, S1 and S2, bringing the total number of prealbumin alleles in Thoroughbred horses to eight. The gene frequencies of these eight alleles in Australian Thoroughbreds are presented. All eight prealbumin types exhibit antiprotease activity and therefore, it is suggested that the name prealbumin (Pr) should be abandoned in favour of protease inhibitor (Pi) although at this st...
Gel electrophoresis of rotavirus RNA derived from six different animal species. Rotavirus RNA prepared from calf, pig, mouse, deer, foal and dog-adapted human isolates was compared using polyacrylamide gel electrophoresis. Reproducible differences in the RNA migration patterns were found between all isolates. There were 11 clearly resolved segments in the pig, mouse and foal samples. The calf rotavirus RNA and deer rotavirus RNA separated into 9 bands and 10 bands, respectively. The dog-adapted human virus migrated in 12 bands, and this probably results from the complex passage history of the original human rotavirus isolate.
Isoenzymes of equine alkaline phosphatase. Alkaline phosphatase isoenzymes from small intestine, cecum, large colon, small colon, liver, kidney, leukocytes, and serum from ten clinically normal horses were defined by their sensitivities to L-phenylalanine, L-homoarginine, levamisole and heat, and by polyacrylamide gel disc electrophoresis. Readily identifiable isoenzymes occurred in small intestine, granulocytes, kidney, cecum, and large and small colon. By contrast, alkaline phosphatases from liver, lymphocytes, and serum could not be discriminated by this group of tests.
Identification of alpha1-lipoproteins in crossed immunoelectrophoresis. Evans Blue dye binds selectively, but with different avidities, to five major antigens in human serum. The anodic mobility of the antigen-dye complexes is greater than that of the antigens alone in crossed immunoelectrophoresis, which is of practical value for identification. We used this characteristic to show that in some human sera there is a population of alpha1-lipoprotein molecules that migrates electrophoretically in the beta-lipoprotein region, where in conventional zone electrophoresis it could be mistaken for beta-lipoprotein. We also demonstrate that horses, unlike rabbits, rarely m...
The effect of binding ions on the oxidation of horse heart ferrocytochrome c. The research explores how different binding ions affect the oxidation speed of horse heart ferrocytochrome c, a protein, by potassium ferricyanide at a constant ionic strength. Studying the Ion Effect […]
The lectin-binding sites of the erythrocyte membrane components of horse, swine and sheep. Characterization by their molecular weights. The membrane components of equine, porcine and ovine erythrocytes were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis and subsequently incubated with the radioiodinated lectins from lentils (LCH), castorbeans (RCA), Phaseolus beans (L-PHA), gorse seeds (UEH-F) and from vineyard snails (HPA). The following individual glycoproteins could be labeled: gp 26, 33, 100 and 320 in horse erythrocytes, gp 24, 46, 75, 130 and 210 in swine and gp 24, 57, 100 and 210 in sheep erythrocytes.
Determination of the charge of horse kidney metallothionein by free boundary electrophoresis. Traditionally, the charge of a protein molecule as determined by electrophoresis has been compared to that revealed by pH titration, and any lack of coincidence has been ascribed to ion binding, and the two results have been brought into agreement by adjustment of binding parameters (1). Metallo-thionein allows a unique opportunity to examine the validity of the electrophoretic approach, since the amino acid sequence and metal atom binding studies allow the absolute charge of the molecule to be computed (2). This then can be compared to the charge determined from electrophoretic mobility measu...
Genetic polymorphism and close linkage of two alpha 1-protease inhibitors in horse serum. Two-dimensional electrophoretic analysis of horse serum proteins was done by a first-dimension separation in agarose gel (pH 5.4) followed by a second-dimension separation in horizontal polyacrylamide gel (pH 9.0). This method resulted in improved and reproducible separation of many alpha-globulins. Two groups of alpha 1-globulins, designated Pi1 and Pi2, were found to be protease inhibitors. Preliminary studies indicated that Pi1 and Pi2 proteins differed from each other in molecular weight and in protease inhibiting spectra. Extensive polymorphism was observed for both these proteins. Family...
Hemoglobin polymorphism in Equus przewalskii and E. caballus analyzed by isoelectric focusing. 1. Through the use of isoelectric focusing and peptide analysis, the hemoglobins of Przewalski's horse. Equus przewalskii and the domestic horse, E. caballus have been compared. 2. Przewalski's horses have two separate alpha-globin chain polymorphisms similar to domestic horses. Each hemoglobin phenotype could be accurately determined by isoelectric focusing. 3. Confirmation of the electrofocusing hemoglobin determinations was made by comparison to amino acid composition analyses of purified tryptic peptides and by analysis of the rare hemoglobins phenotypes observed in a family of Norwegian t...
The characterization of equine prealbumin (Pr) proteins by antigen-antibody crossed electrophoresis. Acta vet. scand. 1979, , 180–190. — Selected equine Pr phenotypes from a total of 55 horses of mixed breeds were investigated. The horse sera were subjected to acid starch gel electrophoresis at pH 4.8, followed by right angle electrophoresis in agarose gels containing rabbit-produced anti-Pr protein. This technique gives peaks in the agarose gels corresponding to the Pr zones in acid gels. The investigation revealed patterns of the Pr protein which were more complex than those seen when using ordinary acid starch gel electrophoresis. The phenotypes FF, II and LL showed a total of eight p...
Lipid composition and cholesterol esterification in serum lipoprotein fraction of the horse, Equus caballus. 1. Changes in lipid components of lipoproteins during incubation of horse serum at 37 degrees C were investigated. In non-incubated serum, cholesterol and lecithin existed predominantly in alpha-lipoprotein or in high-density lipoprotein (HDL). Lysolecithin was mainly associated with the fraction with density above 1.21. 2. When serum was separated into alpha- and beta-lipoproteins by the heparin precipitation method after 1 hr incubation, the decrease in alpha-lipoprotein free cholesterol and lecithin was about four times that in beta-lipoprotein counterparts. 3. When serum lipoproteins were ...
Isoelectric focusing of some enzymes from Echinococcus granulosus (horse and sheep strains) and E. multilocularis. Extracts of the horse and sheep strains of Echinococcus granulosus and E. multilocularis were compared on the basis of their isoenzyme patterns for 10 enzymes by means of isoelectric focusing in polyacrylamide gels. The enzymes examined were: acid phosphatase, lactate dehydrogenase, malate dehydrogenase, malic enzyme, phosphoglucoseisomerase, isocitrate dehydrogenase, adenylate kinase, aldolase and alpha-glycerophosphate dehydrogenase. Interspecific and intraspecific differences are apparent in the isoenzyme profiles of all the enzymes except adenylate kinase; the pattern and activity of adeny...
Analysis of mechanisms regulating the expression of parental alleles at the GPD locus in mule erythrocytes. Erythrocyte glucose-6-phosphate dehydrogenase (G6PD) was examined by 13% starch gel electrophoresis in 74 mules (42 females and 32 males), 35 donkeys, and ten horses. The quantitative expression of the parental alleles at the Gpd locus varies greatly in female mules from the hemizygous expression of the maternal allele to that of the paternal. The data obtained indicate that the X chromosomes are randomly inactivated in females mules. No selective advantage of a cell population with a maternally (or paternally) derived X active was found in female mule erythrocytes. It is suggested that the ph...
Demonstration isolation and identification of culturable microfungi and bacteria in horse hair and dandruff. Immunochemical comparison with allergic components. Horse hiar and dandruff have been investigated for their content of microfungi and bacteria. Inoculation and incubation on V-8 agar containing penicillin and streptomycin, with subsequent colony counting and identification, revealed more than nine and five different genera of microfungi and bacteria respectively, in horse hair and dandruff. Isolation and cultivation of the quantitatively dominating species, and preparation of an extract of these were performed, followed by immunochemical comparison with extract of the horse hair and dandruff using crossed-line immuno-electrophoresis. As no imm...
A new allele in the prealbumin system of horse serum markers. A family study of an index case in the Arabian breed of horses demonstrated the presence of a new allele in the prealbumin (Pr) system of electrophoretically determined markers in horse serum which, when homozygous, results in the absence of any recognizable zones in the Pr region. The symbol PrO is proposed for this allele which has an estimated frequency in Arabian horses of 0.09.
Isoelectric focusing of horse serum esterase isozymes and detection of new phenotypes. A new method for separating the isozymes of horse serum esterase is described. The improved resolution has enabled us to detect several previously undescribed phenotypes. This method has also been used to detect two different apparently 'silent' alleles.
Common and species-specific esterases of Equidae–IV. Horse of przewalski, onager and Zebra hartmannae. 1. Among several species of Equidae only E. przewalskii possesses a serum esterase identical with that of E. caballus. 2. The esterases of Hemionidae differ slightly from that of domestic horse by electrophoretic migration and by antigenic structure. 3. Zebras (grevyi, burchelli) appear devoid of this component but Z. hartmannae possesses an esterase of high enzymatic activity, differing notably from that of horse by electrophoretic and antigenic properties.
Polymorphic post-albumin of cattle and horse plasma identified as vitamin D binding protein (Gc protein). Cattle and horse plasma samples of known post-albumin types were radiolabelled with 14C-vitamin D3. These samples were then analysed by polyacrylamide gel electrophoresis, followed by autoradiography. The patterns observed were identical to those of post-albumin variants. The polymorphic post-albumin protein of cattle and horse was thus identified as the vitamin D binding protein and homologous to the Gc protein of human plasma.
Genetic polymorphism of the vitamin D binding protein and another post-albumin protein in horse serum. Horizontal polyacrylamide gel electrophoreses, on 10% separation gel, of horse serum revealed polymorphism of the vitamin D binding protein (Gc protein) and another post-albumin protein (Pa). Family data supported the hypothesis that Gc and Pa types were controlled by autosomal codominant alleles. For both Gc and Pa proteins, the homozygous types showed a single fraction while the heterozygous type had two fractions. Pa types were found to be identical to the post-albumin types reported earlier by starch gel electrophoresis. Two Gc alleles, GcF and GcS, and three Pa alleles, Pa D, Pa F and Pa ...
Lactoperoxidase-catalyzed iodination of horse cytochrome c:monoiodotyrosyl 74 cytochrome c. Iodination of horse cytochrome c with the lactoperoxidase-hydrogen peroxide-iodide system results initially in the formation of the monoiodotyrosyl 74 derivative. This singly modified protein was obtained in pure form by ion exchange chromatography and preparative column electrophoresis. It shows an intact 695 nm absorption band, the midpoint potential of the native protein, a nuclear magnetic resonance spectrum which indicates an undisturbed heme crevice structure, a normal reaction with antibodies directed against native horse cytochrome c, and circular dichroic spectra in which the only cha...
