Starch Gel Zone Electrophoresis Research Articles

Die diagnostische bedeutung der analyse von serumproteinen durch stärkegelelektrophorese

By zone electrophoresis in starch gel 100 sera from blood donors not related to each other and 430 sera from subjects with various diseases were examined. The incidence of haptoglobin types corresponds with the European incidence. The significance of this auxiliary method for diagnostic purposes resides in the fact that it permits the demonstration of hetergeneous γ-globulin fractions, which in starch gel appear as one or several more intensively stained stripes. They were observed in malignant tumoral processes, in lupus erythematosus disseminatus and in Hodgkin's disease. Cirrhosis of the liver also gives a characteristic picture. Further, paraproteins were divided into fractions, i.e. they were distinguished from macroglobulins. After repeated infusions of native or dried plasma, no changes in the haptoglobin type were observed.

Zymograms of the Parasitic Nematodes, Neoaplectana glaseri and N. carpocapsae, Grown Axenically

The application of a technique for separating different molecules (zone electrophoresis in starch gel) combined with identification techniques (chemically specific staining for enzymatic activity), i.e., the zymogram, was used to show molecular similarities and differences among enzymes of two parasitic nematodes grown axenically. The esterases of Neoaplectana glaseri Steiner, 1929, were multiple and varied in activity to substrates. They were similar for worms grown on media from natural sources or in a medium that was in part chemically defined. They were insensitive to eserine and activated by taurocholate. In addition, they were qualitatively similar for populations of worms composed almost exclusively of third-stage larvae as compared to populations of worms which contained adult and larval stages. The esterases of N. carpocapsae Weiser, 1955, differed from those of N. glaseri. Neither species showed alkaline phosphatase or lactic dehydrogenase activity. Acid phosphatase, present in multiple molecular forms in N. glaseri, differed from that of N. carpocapsae. Working with parasites in culture, one must consider many criteria of adaptation or change. Morphological changes might be the last and least apparent. Subtler criteria such as infectivity, antigenicity, metabolic activity, and genetic transformability may correspond to molecular differences. The quality and/or quantity of these can mirror not only adaptations from nature to culture, or from one type of culture to another, but also differences between related species and changes associated with development. The evidence so far is that Neoaplectana glaseri Steiner, 1929, a nematode of certain insects (Glaser and Fox, 1930), has changed little under axenic conditions compared to protozoa or cultured mammalian cells. No changes in morphology or infectivity have appeared during 17 years or ca. 216 cultured generations (Stoll, 1953, and personal communication). The present work is a study of molecular similarities and differences among selected enzymes of N. glaseri at different stages of development and grown under different axenic conditions. Comparison is made with enzymes of another nematode of insects, Received for publication 19 March 1963. * Present address: Division of Life Sciences, The University of California, Riverside. N. carpocapsae Weiser, 1955. The major tool used, the zymogram, combines a technique for separating molecules according to differences of size and charge (zone electrophoresis in starch gel) and identification techniques (chemically specific staining for enzymatic

Studies of the plasma proteins in pregnancy using starch gel zone electrophoresis

Serum proteins in 126 serum samples from 100 pregnant women were analyzed by starch gel zone electrophoresis. An elevation of the slow α 2-globulin ( α 2-macroglobulin) was noted in 59% of the samples, an increase of β-globulin (transfenin) in 29%, and an increase of fast α 2-globulin and decrease of albumin and γ-globulin in approximately 12%. Samples from only 3 patients showed the unusual finding known as the “pregnancy band” migrating in front of the slow α 2 fraction. Changes of the pattern during the course of pregnancy have been discussed.

Properties and classification of the soluble esterases of human liver

Water-soluble enzymes of human liver were separated by zone electrophoresis in starch gel and compared with those of serum. Three separate zones of esterase activity in the liver extracts were found. An attempt was made to identify these enzymes by using several series of substrates and inhibitors in conjunction with various histochemical staining techniques. One enzyme, migrating at pH 7.2 toward the cathode, reacts like an acetylesterase. Another enzyme, migrating toward the anode at a rate approximating that of serum albumin, reacts like an aliesterase. A third region of activity contains a mixture of esterases and acid phosphatase. The organophosphorus anticholinesterases that were tested showed different patterns of inhibition of the liver esterases. Observations on alkaline phosphatase, sulphatase, glucosidase, galactosidase, and leucine aminopeptidase were recorded for comparison with the data on esterases.

Identification of Fish Species by Starch Gel Zone Electrophoresis of Protein Extracts

Identification of Fish Species by Starch Gel Zone Electrophoresis of Protein Extracts

Dehydrogenases of neurospora crassa.

Malate, isocitrate, glucose-6-phosphate, and 6-phosphogluconate dehydrogenases of the homogenate prepared from the mycelia of several wild strains of Neurospora crassa have been subjected to zone electrophoresis in starch gel. Four electrophoretically different malate dehydrogenases, a single isocitrate dehydrogenase, three glucose-6-phosphate dehydrogenases, and two 6-phosphogluconate dehydrogenases were obtained regardless of the strain or the media in which the organism was grown.

Cell Bound ^|^alpha;-Amylase in Aspergillus oryzae

α-Amylase in Aspergillus oryzae is generally known to be present in mycelium as well as in medium, whether it is grown with or without shaking. The interesting fact is reported in this paper, when A. oryzae was grown in a phosphate deficient medium, however, α-amylase accumulation occurred only within the cell. From the results of pH-activity curve, starch gel zone electrophoresis and differential centrifugation, there seemed to be no distinctions between exo and endocellular α-amylase. The liberation of α-amylase from the cell was observed to be dependent on pH and the concentrations of anions. The significance of the α-amylase accumulation and liberation in this system is discussed.

Multiple forms of esterases from human erythrocytes.

SummaryA system is described in which multiple bands of esterase activity can be resolved from human red cell hemolysates by zone electrophoresis in starch gel. Variations from the normal pattern were observed in certain patients with liver disorders and associated anemias. Another atypical esterase pattern was found in a pair of monozygotic twins and their mother indicating that the observed variation is under genetic control.

Species Identification by Starch Gel Zone Electrophoresis of Protein Extracts. II. Meat and Eggs.

Species Identification by Starch Gel Zone Electrophoresis of Protein Extracts. II. Meat and Eggs.

Development of serum haptoglobins in man.

SummaryTo determine the time of appearance of the haptoglobin serum proteins in the developing human being, starch gel zone electrophoresis was performed on sera from 5 fetuses, 63 umbilical cord bloods, and 86 infants. Haptoglobins were absent in sera from the fetuses, but present in 10% of the umbilical cord bloods at birth, in 75% of the sera from infants up to 2 weeks old, and in 100% of the sera from the infants over 2 weeks of age.For cooperation and assistance in securing the cord and newborn bloods necessary for this study, we wish to thank Dr. Marion Erlandson and Dr. Margaret Hilgartner, Dept. of Pediatric Hematology, New York Hospital, Dr. Roy Bonsnes and Miss Dorothy Wendel, Central Laboratories of New York Hospital, and Dr. Wha Shim Han, Bellevue Hospital. We also wish to thank Dr. Bartholomeus Hoogstraten, Mt. Sinai Hospital, for obtaining the fetal bloods. Dr. German Castillo, Univ. of the Philippines, participated in the early phases of the study.

Starch-gel electrophoresis of anterior pituitary hormones.

SEPARATION of pituitary hormones by zone electrophoresis on paper has not proved very successful, possibly due to complex formation between proteins1. Starch-gel zone electrophoresis introduced by Smithies2,3 separates proteins not only by their net charge but also by their molecular size. This procedure has superior resolving power to paper or agar-gel zone electrophoresis and has been successfully applied to the analysis of thyrotrophic hormone4–6. Starch-gel electrophoretic analyses of various standard preparations of anterior pituitary hormones are presented here together with some evidence of the distribution of hormonal activities.

Localization of prostatic serum acid phosphatase by starch-gel electrophoresis

Serum acid and alkaline phosphatase have been completely separated by zone electrophoresis in starch gel. The prostatic acid phosphatase activity was found in a narrow zone corresponding to the α-β-globulin number 6, the non-specific alkaline phosphatase activity in a zone corresponding to the α-β-globulin number 4.

Separation of human hemoglobins by starch-gel zone electrophoresis

The separation of normal and abnormal human hemoglobins by starch-gel electrophoresis has been described and the relative mobilities of the different components determined. The method of starch-gel electrophoresis has high resolution and was capable of detecting five protein fractions in normal red cell hemolysates. Fetal hemoglobin was readily separable from normal adult hemoglobin; and elevations of the A 2 hemoglobin could be demonstrated in some patients with thalassemia minor although quantitative measurements were not made.

Species Identification by Starch Gel Zone Electrophoresis of Protein Extracts. I. Fish

Species Identification by Starch Gel Zone Electrophoresis of Protein Extracts. I. Fish

Heterogeneity of tissue dehydrogenases

Rat tissue homogenates have been subjected to zone electrophoresis in starch gel. Lactate, malate, isocitrate, glucose 6-phosphate and α-glycerophosphate dehydrogenase activity was directly visualized by incubating the gel strips in solutions containing a specific substrate, a tetrazolium salt, methylene blue and the appropriate pyridine nucleotide. Electrophoretic heterogeneity of some of the dehydrogenases was observed in material from different organs. A similar multiplicity of components in some specific dehydrogenases was also demonstrated in cytologically homogeneous material including cells grown by tissue culture techniques. Significance of this multiplicity of enzymes is discussed.

Frequencies of the Haptoglobin Groups in 406 French Blood Donors

The method of zone electrophoresis in starch gel enabled Smithies1 to describe three haptoglobin groups in human sera. Using a standard technique previously reported2,3 we have examined the sera of 406 blood donors living in Paris. In each ‘Plexiglas’ tray (internal dimensions 234 mm. × 80 mm. × 6 mm.) three serum samples mixed with a haemoglobin solution (0.05 ml. of a solution containing 50 mgm. of haemoglobin being added to 1 ml. of serum) were allowed to migrate simultaneously, side by side, for 18 hr., under a potential of 100 V. After electrophoresis the starch gels were divided into two slices and stained, one with amido black, and the other with benzidine reagent for peroxidase activity. The technique was the same as that previously described and used to detect haemoglobins in agar gels4 but without using zinc acetate as a solution.

Localization of Hæmocyanin on Starch Gel Electrophoretic Patterns

USING the Smithies technique1 of zone electro-phoresis in starch gel, Woods et al. 2 have recently demonstrated the occurrence of several haemocyanin proteins in the blood sera of certain crustacean species. The identification is tentatively based on the occurrence of two or more protein bands of a similar order of mobility and which are extremely concentrated in comparison to the other blood protein components.

澱粉gel中の電気泳動法による Haptoglobin 型の研究

The author examined the haptoglobin (Hp) type of Japanese by means of Smithies' zone electrophoresis in starch gel. The results are as follows:1) In 232 normal adults, 24 showed Hp 1-1 type, 87 Hp 2-1 type and 121 Hp 2-2 type. The gene frequencies are calculated to be Hp1: 0.291 and Hp2: 0.709. The Hp1 frequency of Caucasian and African races previously reported is not lower than one of the Japanese encountered in this study.2) No relation is revealed between the Hp types and the sex, pregnancy, ABO or Rh blood types and serum protein concentration.3) No determinable Hp pattern is detected in cord bloods or bloods of newborn within 5 days of birth.4) By transfusions of the plasma of different Hp types no causal reaction seems to be provoked. In massive transfusions Hp patterns of recipients are influenced by donors' Hp types and show the Hp patterns like those given by the mixed sera in vitro. The modified patterns disappear in about 2 days. The relative rapid disappearance of transfused Hp may be chiefly due to increased haemolysis in vivo.

Zone Electrophoresis in Starch Gels and its Application to Studies of Serum Proteins

Publisher Summary This chapter reviews that the starch-gel electrophoretic method can be applied to the investigation of many complex protein mixtures in addition to serum proteins, for example, enzymes, hormones, and tissue extracts. One aim of this chapter is to describe the current technical problems associated with starch-gel electrophoresis and to consider their possible solutions. A second aim is to summarize the information already obtained by application of the method to studies of serum proteins and their inheritance, and to put this information into perspective in relation to future work. The chapter discusses the experimental procedures used for starch-gel electrophoresis and focus on the technical details is important for the realization of the best starch-gel separations. It provides a summary of the data relevant to the specific identification of serum components demonstrated by starch-gel electrophoresis. The chapter also reviews the results and significance of recent studies of serum proteins using the method. The nature of the physical factors governing the electrophoretic separations obtained in starch gels is discussed. The hypothesis is considered that molecular size plays a particularly important role in the separation processes, and the evidence in support of this is presented. Evidence concerning the validity of the results obtained in starch gels is also reviewed.

Zone electrophoresis in starch gels; a report on Smithies' method in normal adults and in patients with tuberculosis.

Zone electrophoresis in starch gels; a report on Smithies' method in normal adults and in patients with tuberculosis.