Solvent extraction of serum and other biological fluids at an acidic pH is a convenient method to remove the insulin-like growth factor binding proteins (IGFBPs); however, an incomplete removal of IGFBPs can occur and this can potentially interfere with the radioimmunoassay of insulin-like growth factors (IGFs). This study compared the ...

We purified human procollagen type I carboxyl-terminal propeptide (PICP) that had been cleaved as in vivo from procollagen. PICP in serum-free medium from cultured human fetal fibroblasts was purified by thiophilic adsorption chromatography, low-pressure gel filtration, and HPLC gel filtration. The purity and homogeneity of the protein was verified by ...

A monoclonal antibody was developed to the 20 kd major allergen of cow by immunizing mice with crude dander extract. The monoclonal antibody did not exhibit cross-reactivity to cat, dog, and horse dander extracts when studied by ELISA inhibition. The antibody was used in affinity chromatography for the purification of ...

A cationic (pI 8.3) wall-bound peroxidase has been purified to homogeneity from suspension-cultured cells of French bean (Phaseolus vulgaris L.). The enzyme was a glycoprotein and its M(r) was 46,000 as determined by SDS/Page and h.p.l.c. gel filtration. It was localized biochemically to microsomes and the cell wall, and the ...

Two vitellins (Vn-1 and Vn-2) were identified in cockroach ovary. Vn-2 was purified by ion-exchange chromatography using DEAE cellulose and Sepharose CL-6B, and gel filtration using Sephadex G-200 and Sepharose 4B. Vn-2 comprises three subunits with molecular weights of 102, 72 and 40 kDa. Vn-2 contains diacylglycerol and triacylglycerol; their ...

The yeast Saccharomyces cerevisiae was entrapped within polyacrylamide gel beads by employing a procedure that uses sodium dodecylsulfate as a detergent to improve the spherical configuration of the beads. The resulting preparation showed a rate of fumarate bio-conversion to L-malic acid about 60 times higher than that found for the ...

The small protein ubiquitin (76 amino acids) has been synthesized under optimized conditions by Merrifield solid-phase methodology using the N alpha-Fmoc protecting group. The crude polypeptide mixture was purified to homogeneity by gel filtration, dialysis and a combination of cation- and anion-exchange chromatography to yield ubiquitin. Amino acid analysis, enzymic ...

A superoxide dismutase (SOD) from adult worms of Dirofilaria immitis was purified using ethanol-chloroform and acetone treatment, DE 52 cellulose, and Sephadex G-75 gel chromatography to obtain a 677-fold purification and a specific activity of 5483 units/mg of protein. The purified SOD was essentially homogeneous as judged by polyacrylamide gel ...

Soluble fractions and particulate extracts from human prostate, and extracts from rat-liver membranes were used as a source of kinases to phosphorylate endogenous proteins in the presence of gamma- 32P-labeled ATP. Histone was also added as a substrate in order to compare the direct partial acid hydrolysis of phosphoproteins in ...

We have previously reported the evidence for presence of a humoral factor 'injurin', which induces expression of the hepatocyte growth factor (HGF) gene in MRC-5 human embryonic lung fibroblasts. We have now purified a factor from porcine liver which stimulates HGF production but differs from injurin. When injurin activity was ...

A modified procedure for the two-dimensional electrophoretic analysis of basic polypeptides is described. This method uses isoelectric focusing with carrier ampholytes in the first dimension, and sodium dodecyl sulfate-electrophoresis in the second dimension. Counteraction of the cathodic drift is achieved by glass tube treatment (silanization), electrolyte modification (use of weak ...

A cyclophilin type peptidyl-prolyl cis-trans isomerase was purified from a halophilic archaeum, Halobacterium cutirubrum DSM 669. The activity increased with an increase in KCl concentration up to 4 M. Sensitivity to cyclosporin A was comparable to that of eukaryotic cyclophilin and was also affected by KCl. IC50 for Cyclosporin A ...

A Kunitz type proteinase inhibitor was isolated from extracts of the sea anemone Stichodactyla helianthus. The purification procedure comprises treatment with trichloroacetic acid followed by affinity chromatography on trypsin-Sepharose and gel filtration on Sephadex G-50 or ion exchange chromatography on CM-cellulose. The major inhibitor (isoelectric point 8.4) is a small ...

In this study two phosphopeptide (PP) fractions from purified, calf thymus nuclei, prepared under protective conditions were characterized. The nuclei were lysed and dialyzed, and the material in the dialysates was fractionated by anion exchange column chromatography. This revealed the presence of two main phosphopeptide (PP) fractions, numbers 1 and ...

1. Phospholipase A2 was isolated from the venom of Agkistrodon bilineatus by Sephadex G-75 and CM-Cellulose column chromatographies. 2. The purified phospholipase A2 gave a single band on disc polyacrylamide gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis and ODS-HPLC. 3. The enzyme preparation had a mol. wt of 14,000, ...

An extract from intact spores of Bacillus cereus T having a germination-inducing activity was studied. Two distinct germinative principles were found through dialysis of the extract. One was diffusible through the dialysis membrane and the other was non-diffusible. The activity of the former fraction was inhibited by the addition of ...

We have previously demonstrated that Corynebacterium cyclohexanicum degrades cyclohexanecarboxylic acid, a bacteriocide, through a pathway including the aromatization of 4-oxocyclohexanecarboxylic acid to 4-hydroxybenzoic acid [Kaneda, T. (1974) Biochem. Biophys. Res. Commun. 58, 140-144]. Aromatization has now been shown to be catalysed by two desaturase enzymes. Under the action of desaturase ...

A rapid two-step procedure has been developed for the purification of Despro(2)-Val15-Leu17-aprotinin from the culture supernatant of a recombinant yeast by affinity and ion-exchange chromatography. DesPro(2)-Val15-Leu17-aprotinin was purified to homogeneity, as demonstrated by dodecylsulfate gel electrophoresis and analysis of the N-terminal amino acid sequence. (c) 1993 John Wiley & Sons, ...

An enzyme fraction with catalytic activities for the biosynthesis of the pipecolic acid containing cyclopeptolide SDZ 90-215 was partially purified and characterized from the genus Septoria. The crude cell homogenate was subjected to polyethyleneimine precipitation, ammonium sulfate precipitation and FPLC gel filtration. The denatured enzyme shows an apparent molecular mass ...

Motilin was isolated from cat small intestine by a series of chromatographic steps. Using a radioreceptor assay, based upon binding of iodinated porcine motilin to rabbit antral smooth muscle membranes, it was shown that cat duodenal mucosa contains about 495 ng/g tissue, the jejunal mucosa 161 ng/g tissue and the ...

Two types of affinity gels consisting of cross-linked polyacrylamide and affinity ligands possessing nucleic acid bases were prepared. One type of gel was polyacrylamide-poly(vinylnucleobase) conjugated gel, where the poly(vinylnucleobase) such as poly(9-vinyladenine) (PVAd) bearing a nucleobase in the side-chain was entrapped in the gel matrix. The other type of gel, ...

Fractions obtained by gel filtration or ultrafiltration of dog serum were tested for their mitogenic activity on canine prostatic epithelial cells: two prostatic growth factor (PGF) entities were found, a major one of 150 kDa (PGF-I) and a minor one of 1.5-2.0 kDa (PGF-II). Treatment and/or extraction with acetic acid, ...

The clotting enzyme (Stenoxobin), from the venom of Lachesis muta stenophyrs, was purified by gel chromatography on Bio-gel P-100 followed by agmatine CH-Sepharose-4B and FPLC on Mono Q column. By SDS polyacrylamide gel electrophoresis the mol. wt was found to be 37,000. The enzyme is a glycoprotein with 1.6 moles ...

Degraded macromolecular lignin, which was isolated from the effluents of commercial pulp processing and known to inhibit early development in marine organisms, was separated and characterized using several polyacrylamide gel electrophoresis (PAGE) techniques. This lignin-derived macromolecule (LDM), when subjected to native PAGE and stained with alcian blue, appeared as a ...

Two forms of acidic arginine amidases were separated from human kidney extract using the techniques of basic ion exchange adsorption and elution as well as lima bean trypsin inhibitor (LBTI) and aprotinin affinity adsorptions and elutions. The enzymes were tentatively named acidic human renal arginine amidase-L (AHRAA-L, with affinity to ...

A relationship between plasma membrane fatty acid binding protein (FABPpm), a putative membrane transporter for long-chain fatty acids, and the mitochondrial isoform of aspartate aminotransferase (m-AspAT) has been reported. Accordingly, we have compared the chemical and immunological properties of rat liver m-AspAT with those of rat liver FABPpm isolated by ...

Borrelia hermsii, Borrelia parkeri, Borrelia anserina, and Borrelia coriaceae produced several lipoproteins identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of bacteria grown in [3H]palmitate. Five major acylated proteins were demonstrated by sequential alkaline and acid hydrolysis. High-pressure liquid chromatography of isolated proteins confirmed that covalently bound radioactivity was ...

Alginate gel beads containing indomethacin (IMC) were prepared using a gelation of alginate with calcium cation. Though IMC was not released from the beads at pH 1.2 because of its extremely low solubility in aqueous media, it was released according to the swelling of alginate gel at pH 6.8. The ...

The N-glycolylneuraminic acid-specific lectin (AFL) from the foot muscles of the marine clam Anadara granosa has been purified to homogeneity by affinity chromatography on bovine submaxillary mucin-Sepharose 4B. The Ca(2+)-dependent lectin agglutinates rabbit erythrocytes. The purified lectin is a tetrameric protein of native M(r) 254 kDa having a pI value ...

Base-specific separations of oligodeoxynucleotides were achieved with high resolution by electrophoresis, using a urea-gel capillary, in which poly(9-vinyladenine) (PVAd) was utilized as an affinity ligand. The migration behaviour and the plate number of oligodeoxynucleotides were investigated as a function of urea concentration between 2 and 10 M in capillary gel ...

Analysis of amino acids is complicated by treatment with ampicillin. High voltage electrophoresis, which is convenient for the qualitative assessment of metabolic diseases, yields smears of ampicillin that mask the bands of citrulline, homocitrulline, phenylalanine, cystine, and homocystine. The addition of penicillinase prior to high voltage electrophoresis eliminates ampicillin and ...

BACKGROUND: Cyn d I has been found to be the major allergen of Bermuda grass (Cynodon dactylon) pollen, but its exact nature remains to be clarified. METHODS: Cyn d I, the major allergen of Bermuda grass (Cynodon dactylon) pollen, was purified by monoclonal antibody (MoAb) affinity chromatography, and its biochemical ...

Bovine myeloperoxidase (MPO) was isolated and purified from neutrophil granules using protein extraction at pH 4 and gel filtration combined with fast protein liquid chromatography. The extracted protein was identified as MPO based on its absorption spectrum, amino acid composition, peroxidase activity and polypeptide structure. Bovine neutrophils contained three different ...

Isoforms of phospholipase A2 (PLA2) from the venom of Naja mossambica mossambica (the spitting cobra) were purified by a combination of gel filtration on Bio-gel P-30 and ion exchange chromatography on DE-52 Cellulose and the purification followed by three types of polyacrylamide gel electrophoresis. SDS PAGE failed to resolve the ...

The fractionation of Phoneutria nigriventer spider venom by gel filtration (Sephadex G-10-120) followed by ion-exchange chromatography (microgranular CM-cellulose-52) resulted in sixteen fractions (CI to CXVI) from which CVII+VIII, CIX and CX+XI caused dose-dependent and short-lived contractions of both arterial and venous rabbit vessels. Fraction CX+XI was further purified by a ...

Two forms of highly purified prolactin (PRL) were obtained from pituitary glands of Xenopus laevis by extraction of acetone-dried powder with acid acetone and high-performance liquid chromatography on anion exchange, gel filtration, and reverse-phase columns. Purification was monitored by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis employing antiserum against ...

Agarose gel electrophoresis is a powerful technique for the separation of nucleic acids on the basis of their size and conformation. The development of methods to recover size-fractionated nucleic acids molecules from agarose gels has greatly facilitated recombinant DNA technologies. Although several methods for recovering DNA and RNA molecules have ...

Phospholipase D (phosphatidylcholine phosphatidohydrolase, EC 3.1.4.4) was purified from cabbage leaves. The two step purification procedure involved hydrophobic chromatography on Octyl-Sepharose followed by a Mono-Q/FPLC-column with a total yield of 23% and a purification factor of 1000. A zymographic assay was used to detection of PL D activities at various ...

Biosynthesis of the vicinal diol fatty acid (12R,13S)-dihydroxy-(5Z,8Z,10E,14Z)-eicosatetrae noic acid from arachidonic acid was studied in preparations of the red alga Gracilariopsis lemaneiformis. The transformation consisted of initial 12-lipoxygenase-catalyzed oxygenation of arachidonic acid into (12S)-hydroperoxy-(5Z,8Z,10E,14Z)-eicosatetraeno ic acid followed by hydroperoxide isomerase-catalyzed conversion of the hydroperoxide into (12R,13S)-dihydroxyeicosatetraenoic acid. Short time ...

Because previous purification procedures for human kappa-casein may have caused the loss of some carbohydrate, relatively gentle methods were used. The protein was isolated by a four-step procedure which included isoelectric precipitation of whole casein, gel chromatography on Sephadex G-200 in the presence of SDS, removal of the SDS with ...

A lectin was isolated from hemolymph of the Southeast Asian horseshoe crab Tachypleus gigas by using glycophorin HA affinity chromatography and Sephacryl S-300 gel filtration. The purified lectin had a molecular mass of approximately 396kDa and was composed of 13 identical subunits with molecular masses of 31 kDa. The serological ...

The composition and antacid properties of six samples of codried antacids containing both aluminum and magnesium were determined. Aluminum hydroxide-magnesium carbonate codried gel and aluminum hydroxide-magnesium hydroxide codried gel were non-homogeneous, as the samples contained combinations of hydrotalcite, amorphous aluminum hydroxide, magnesium hydroxide, magnesium hydroxycarbonate, and magnesium carbonate. All samples ...

Two analytical methods for human secretory-type ribonuclease, which are based on polycytidylic acid/ethidium bromide fluorescence, have been developed. The first is a method for measurement of secretory-type ribonuclease activity utilizing the radial diffusion of ribonuclease in a thin agarose gel plate containing polycytidylic acid and ethidium bromide. Ribonuclease activity was ...

An angiogenic factor from human transitional cell cancer of bladder has been purified by protein extraction, cation exchange chromatography, gel filtration high-performance liquid chromatography (GE-HPLC) and reversed-phase high-performance liquid chromatography (RP-HPLC). The purified substance was named bladder cancer angiogenic factor (BCAF). Biological activity of the BCAF was assessed by the ...

Stimulation of lamb tracheal smooth muscle fibre strips with endothelin-1 and okadaic acid results in the development of isometric tensions which are long lasting. However, endothelin-1 is more potent constrictor than okadaic acid. An analysis of 20,000 Da regulatory myosin light chain by two-dimensional gel electrophoresis shows an identical phosphorylation ...

The potent cytotoxin of Clostridium difficile, toxin B, is internalized by endocytosis and activated intracellularly by an unidentified mechanism. Here it is shown that dithiothreitol treatment of toxin B resulted in (1) a mol. wt of 250,000 which is the smallest species of this toxin shown to be cytotoxic; (2) ...

Two electrophoretic techniques have been developed for the analysis of synthetic beta-A4 related peptides. The first is an acetic acid--urea--polyacrylamide gel electrophoresis denaturing system while the second employs capillary zone electrophoresis at pH 7. Both methods were calibrated to identify the state (monomeric or aggregate) of the longer amyloid fragments ...

In a previous study we reported the presence in human follicular fluid (hFF) of a FSH receptor-binding inhibitor (hFSH-BI) with FSH agonist activity, which was immunologically similar to FSH but could be distinguished from FSH on the basis of its greater stability in acid. We have now purified hFF-derived hFSH-BI ...

Avocado (Persea americana) fruit experience a rapid and extensive loss of firmness during ripening. In this study, we examined whether the chelator solubility and molecular weight of avocado polyuronides paralleled the accumulation of polygalacturonase (PG) activity and loss in fruit firmness. Polyuronides were derived from ethanolic precipitates of avocado mesocarp ...

The N-glycans have been removed by peptide-N-glycosidase F (PNGase F) from purified human non-secretory RNases derived from kidney, liver and spleen. The spleen RNase was purified by two procedures, one of which did not include the usual acid treatment step (0.25 M H2SO4, 45 min, 4 degrees C), to determine ...