Electrophoretic and immunological charactorization of rat neurophysin

1. The electrophoretic properties of rat posterior pituitary proteins have been compared on starch gel with those of bovine and porcine neurophysins. 2. [35S]-Cysteine was injected into the supraoptic nucleus of male rats and 16–24h later the distribution of labelled neural-lobe protein in starch and polyacrylamide gels was determined. In both systems a single major protein component was found to contain more than 80% of the total recovered radioactivity. Between 5 and 10% of the radioactivity was found in a minor component in polyacrylamide gel. 3. In agar, microimmuno-diffusion and -electrophoresis of the rat neural-lobe proteins gave a single arc with neurophysin antiserum, and after starch-gel electrophoresis this arc was shown to be due to the major labelled component. 4. The molecular weights of the rat neural-lobe proteins were estimated by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The molecular weight of the major labelled component was found to be 12000. 5. It is concluded that the rat neurophysin consists of one major and possibly one minor component.

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Purification, properties and specificity of the restriction endonuclease from Bacillus stearothermophilus

Biochemical Journal ◽

10.1042/bj1770049 ◽

1979 ◽

Vol 177 (1) ◽

pp. 49-62 ◽

Cited By ~ 27

Author(s):

C M Clarke ◽

B S Hartley

Keyword(s):

Enzyme Activity ◽

Restriction Endonuclease ◽

Dna Cleavage ◽

Gel Electrophoresis ◽

Bacillus Stearothermophilus ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Major Protein ◽

Activity Restriction

The restriction endonuclease BstI was purified from 70kg of Bacillus stearothermophilus. The final product is at least 97% pure as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; this major protein species co-migrates with the enzyme activity on native polyacrylamide-gel electrophoresis and isoelectric focusing. Pure restriction endonuclease BstI has a subunit mol.wt. of 26,000 and is probably a loosely associated dimer. The enzyme shows maximum activity at pH values between 7 and 9.5, and in the presence of 0.5-2mM-Mg2+. NaCl inhibits the restriction enzyme activity. Restriction endonuclease BstI cleaves DNA in a position identical with that cleaved by endonuclease BamHI (for Bacillus amyloliquefaciens), i.e.: (formula: see text). In the presence of high concentrations of enzyme, DNA cleavage occurs at secondary sites. This side-specificity is enhanced by the addition of glycerol. Preliminary studies indicate that these sites are of the type: (formula: see text).

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STUDIES OF A THYROID HORMONE AND ANDROGEN DEPENDENT PROTEIN IN RAT LIVER CYTOSOL

Acta Endocrinologica ◽

10.1530/acta.0.0840548 ◽

1977 ◽

Vol 84 (3) ◽

pp. 548-558 ◽

Cited By ~ 4

Author(s):

Wolfgang H. Dillmann ◽

Enrique Silva ◽

Martin I. Surks ◽

Jack H. Oppenheimer

Keyword(s):

Molecular Weight ◽

Thyroid Hormone ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Female Rats ◽

Male Rats ◽

Male Rat ◽

Liver Cytosol

ABSTRACT Sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis of the liver cytosol of euthyroid male rats revealed a prominent band (molecular weight, 26 000 daltons), designated Protein II, which was virtually absent in the cytosol of hypothyroid animals. Injection of 500 μg triiodothyronine (T3) per 100 g body weight resulted in a maximal increase in the level of Protein II, reaching 90% of the euthyroidal level 3 days after hormone administration. Concomitant studies with the liver mitochondrial enzyme alpha-glycerophosphate dehydrogenase (α-GPD) indicated that this T3 dose also resulted in a maximal enzyme response in this time period. Since we have estimated that 500 μg of T3 will saturate nearly all nuclear T3 binding sites, these results support the concept that the synthesis of both proteins is limited by nuclear binding. Protein II was absent in the liver cytosol of female rats but could be induced in ovariectomized female rats by androgens. Treatment of male rats with oestradiol resulted in disappearance of Protein II. Since administration of testosterone to hypothyroid male rats caused only a minimal increase in the amount of Protein II, the absence of the protein in hypothyroid animals was not due to androgen deficiency. Similarities in the molecular weight and the response to hormonal manipulation of Protein II and of the urinary α2uglobulin, previously reported by Roy (1973) raise the possibility that these proteins are the same. The high concentration of Protein II in male rat cytosol and the relative ease in its identification by SDS polyacrylamide gel electrophoresis make it a potentially useful model protein for the study of thyroid hormone action at the cellular level.

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Sodium dodecyl sulfate polyacrylamide gel electrophoresis of serum after protein denaturation in the presence or absence of 2-mercaptoethanol.

Clinical Chemistry ◽

10.1093/clinchem/30.3.475 ◽

1984 ◽

Vol 30 (3) ◽

pp. 475-479 ◽

Cited By ~ 26

Author(s):

T Marshall

Keyword(s):

Sodium Dodecyl Sulfate ◽

Gel Electrophoresis ◽

Serum Proteins ◽

Protein Denaturation ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Major Protein ◽

Dodecyl Sulfate ◽

Electrophoretic Techniques

Abstract Human serum proteins were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis after protein denaturation in the presence or absence of 2-mercaptoethanol. Both electrophoretic techniques give characteristic and reproducible banding patterns and achieve a high degree of resolution within the limits of a one-dimensional separation. The major protein bands have been identified, and the merits of the two techniques are compared. Protein denaturation in the absence of 2-mercaptoethanol gives more discrete bands for the purpose of protein identification by maintaining the disulfide-bridge-dependent polypeptide associations characteristic of many serum proteins. However, simultaneous use of both electrophoretic techniques enhances identification by exploiting the response of an individual protein to mercaptoethanol treatment. The clinical potential of this approach for detecting protein disorders is discussed.

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Isolation and protein composition of gap junctions from rabbit hearts

Biochemical Journal ◽

10.1042/bj2050189 ◽

1982 ◽

Vol 205 (1) ◽

pp. 189-194 ◽

Cited By ~ 31

Author(s):

C K Manjunath ◽

G E Goings ◽

E Page

Keyword(s):

Gap Junctions ◽

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Protein Composition ◽

Polyacrylamide Gel ◽

Major Protein ◽

Electron Microscopic ◽

Gap Junctional

We have modified a method for isolating gap-junctional membrane from mouse hearts [Kensler & Goodenough (1980) J. Cell Biol. 86, 755-764] to isolate gap junctions of comparable purity from rabbit hearts more rapidly, with better yield, and without resort to non-ionic detergents. Purification was monitored by electron microscopy of thin-sectioned membrane pellets and by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Gap junctions were obtained as vesicles whose mean surface area approximated that of junctions in intact myocardial cells. About 10-20% of the vesicles were ferritin-impermeable. Approx. 125 micrograms of membrane protein was obtained per 8 g of rabbit heart. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of purified gap junctions showed five major protein bands of mol.wts. 46 000, 44 000, 33 000, 30 000 and 28 500 that co-purified with the junctions. This protein composition was nearly identical with that published for gap junctions of mouse hearts, and differed markedly from the protein composition of gap junctions from non-excitable cells (lens and liver). The constancy of junctional protein composition between hearts of two different species and its non-identity with that from liver and lens suggest that, although gap-junctional structure in mammalian tissues seems to be remarkably similar by electron-microscopic techniques, junctional-channel protein composition actually varies from tissue to tissue and may be adapted to the permeability requirements of the tissue.

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Isoenzymatic pattern of glucose 6-phosphate dehydrogenase from Ascaris suum

Journal of Helminthology ◽

10.1017/s0022149x0001316x ◽

1993 ◽

Vol 67 (3) ◽

pp. 226-232 ◽

Cited By ~ 1

Author(s):

C. Cutillas ◽

B. Rodriguez ◽

P. German ◽

D. Guevara

Keyword(s):

Gel Electrophoresis ◽

Ascaris Suum ◽

Polyacrylamide Gel Electrophoresis ◽

Polyacrylamide Gel ◽

Starch Gel Electrophoresis ◽

Specific Staining ◽

Electrophoretic Patterns ◽

Starch Gel ◽

Glucose 6 Phosphate Dehydrogenase ◽

Isoenzymatic Pattern

AbstractThe isoenzymatic pattern of glucose 6-phosphate dehydrogenase (abbreviation G6PD) from Ascaris suum has been studied by vertical polyacrylamide gel (PAGE) and horizontal starch gel electrophoresis. After polyacrylamide gel electrophoresis, two stained zones could be identified. One corresponded to tetrazolium oxidase activity; since this zone was stained even in the absence of glucose 6-phosphate (G6P), non-specific staining could be detected. In the other zone of activity, seven regularly-spaced bands were identified by staining in the presence of G6P and NADP as substrates. By using starch gel electrophoresis, different electrophoretic patterns for G6PD have been observed in the muscular sac, intestine and reproductive system from A. suum. The existence of three different alleles of G6PD in the same individual suggests the existence of at least two genes for this enzyme.

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Purification and properties of pyruvate kinase from the hepatopancreas of Carcinus maenas

Biochemical Journal ◽

10.1042/bj1530127 ◽

1976 ◽

Vol 153 (1) ◽

pp. 127-134 ◽

Cited By ~ 16

Author(s):

I G Giles ◽

P C Poat ◽

K A Munday

Keyword(s):

Pyruvate Kinase ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Carcinus Maenas ◽

Deae Cellulose ◽

Protein Band ◽

Polyacrylamide Gel ◽

Major Protein ◽

Major Protein Band

Pyruvatekinase from the hepatopancreas of the common shore crab, Carcinus maenas, was purified to a specific activity of 240 units/mg of protein in the assay conditions described. 2. In one method of purification the enzymic activity could be resolved into two fractions after chromatography on DEAE-cellulose. Fructose 1, 6-diphosphate was able to effect the conversion of one form (peak 1) into the second (peak 2). 3. In the presence of a saturating concentration of fructose 1, 6-diphosphate both forms of the enzyme were kinetically similar. 4. Polyacrylamide-gel electrophoresis of the enzyme 1 day after preparation showed a single protein band. On storage at least three protein bands became visible, all of which were associated with pyruvate kinase activity. 5. Chromatography of the enzyme on Sephadex G-200 indicated a mol.wt. of 247000, but in the presence of fructose 1, 6-diphosphate the elution volume of the enzyme increased corresponding to a mol.wt. of 193000. 6 Dissociation of the enzyme in sodium dodecyl sulphate and 2-mercaptoethanol followed by polyacrylamide-gel electrophoresis produced one major protein band with a mol.wt. of 55000.

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Identifikasi Daging Tikus Pada Produk Baso Dengan Metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)

YARSI Medical Journal ◽

10.33476/jky.v26i2.394 ◽

2018 ◽

Vol 26 (2) ◽

pp. 058

Author(s):

Anna P. Roswiem ◽

Triayu Septiani

Keyword(s):

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Sds Page

Bahan baku untuk membuat baso adalah daging hewan, pada umumnya dari daging sapi, ayam, ikan dan babi. Di beberapa daerah di Indonesia terjadi kasus baso tikus. Tujuan penelitian ini adalah menguji ada tidaknya kandungan daging tikus pada produk baso yang dijual di pasar Cempaka Putih-Kecamatan Kramat Jakarta Pusat dan di pedagang baso atau mie baso di sekitar kampus Universitas YARSI Jakarta. Daging adalah protein salah satu metode untuk mengidentifikasi protein adalah metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Hasil penelitian menunjukkan bahwa dari 6 sampel baso terindikasi ada 2 sampel baso dengan nomor 1 dan 5 yang dibuat dari campuran daging sapi dan tikus; ada 1 sampel baso dengan nomor 6 yang terbuat dari daging tikus; dan 2 sampel baso dengan nomor 2 dan 3 yang terbuat dari campuran sapi dan babi, dan hanya 1 sampel baso dengan nomor sampel 4 yang benar-benar terbuat dari daging sapi.

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