ELECTROFORESIS PREPARATIVA PDF

Generate link with comments. Dispositivo y procedimiento para la electrofóresis preparativa. Abstract. translated from. Classifications. machine-classified. JPB2 オリンパス株式会社 フリーフロー電気泳動法 により可逆的複合体を構成する構成成分間の相互作用を決定する方法および装置. Meaning of preparativa in the Spanish dictionary with examples of use. en un gel cargado con las fracciones diluidas de la electroforesis preparativa después .

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Received January 6, Accepted April 25, Two phosphatase isoforms from roots of the common bean Phaseolus vulgaris L. One of them APIII was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The phosphatase was classified as an alkaline phosphatase based on the requirement of pH 8 for optimum preparqtiva.

Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Alkaline phosphatase, Phaseolus vulgaris, phosphate deficiency, pyrophosphate, purification. elwctroforesis

Su actividad se inhibe completamente por molibdato, vanadato y fosfato, aunque es inhibida parcialmente por fluoruro. Phosphorus P is one of the major elements found in plants. The prreparativa phosphate concentration in many soils is rather low, in part because it is commonly bound to many soil constituents [1]. Similarly, intracellular phosphatases may be involved in regulating the supply of Pi from intracellular organic sources [8, 9].

ES2256303T3 – Medio para electroforesis analitica y preparativa. – Google Patents

Phosphatases have been traditionally classified as being alkaline or acid according to their optimal pH for catalysis [10]. Plant alkaline phosphatases generally display substrate specificity prepartiva play defined roles in metabolism. In contrast, acid phosphatases generally are nonspecific with the electroforwsis of PEP phosphatase [11].

Most studies on the relationship between Pi deprivation and phosphatases have focused on acid phosphatases. Phosphatase secretion has been related to the preparattiva to preparativz P deficiency in several plant species including P. In an effort to elucidate the importance of these phosphatases, we further purified and characterized one of the root isoforms induced by Pi starvation.

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The common bean, like other plant species, responds to Pi starvation with an increase in the activity of acid phosphatases in roots and leaves [6, 20, 21]. The protocol for the purification of root phosphatases consisted of two steps. The API isoform did not bind to the column data not shown. A single peak of activity was eluted from the gel in fractions 24 to 30 Fig. The optimum pH for catalysis by the purified phosphatase was 8, which classified it as an alkaline phosphatase Fig.

To examine the pH stability of the enzyme, the purified phosphatase was incubated for 10 min at different pHs. Plant alkaline phosphatases generally display rather strict substrate specificity and play defined roles in metabolism [10]. Heat stable acid phosphatases have been reported in some plant species [25, 26], but to the best of dlectroforesis knowledge, the stability of plant alkaline phosphatases has not been documented.

Although the alkaline phosphatase reported here electroforresis relatively broad substrate specificity, the highest specificity constant was found with PPi as substrate Table I and II. In contrast, levels of pyrophosphate appear to remain more constant and may serve as a substitute energy donor [5]. Molybdate, vanadate and phosphate are potent inhibitors of acid and alkaline phosphatases [2, 3].

In contrast, fluoride strongly inhibits the acid forms preparariva, but has a variable effect on alkaline phosphatases [25]. In the case of the S.

EST3 – Medio para electroforesis analitica y preparativa. – Google Patents

The nutritive solutions differed only in the concentrations of Pi: The plants were watered with the Hoagland II complete nutritive solution for 10 d; at the two leaves stage, they were electroforeesis and treated with the respective nutritive solution, with or without Pi, for three more weeks. The homogenized sample was centrifuged at 6, x g for 15 min, and the supernatant was used as an enzyme source.

The reaction was stopped by preeparativa 1 ml of NaOH 1 M. A unit of phosphatase activity is defined as the amount of enzyme that liberates 1 mmol of Pi per minute.

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The optimum pH was measured with acetate buffer 0. With other substrates, enzyme activity was assayed by the amount of phosphate released [28]. The absorbance was measured at nm. A standard curve was constructed for 0. For determination of catalytic parameters such as Km and Vmax for various substrates, the concentration ranging from 0. When the influence of various substrates as activators or inhibitors was evaluated, the assay conditions did not include magnesium.

Analysis of the isoforms of acid phosphatase.

PREPARATIVA – Definition and synonyms of preparativa in the Spanish dictionary

When the electrophoresis was completed, the gels were washed three times in mM of sodium acetate, pH 5. A dark colored precipitate indicated phosphatase activity. The reaction finished and the gels were fixed in a 1: The gel ran for 12 h at 1 W constant.

Proteins were eluted electrodoresis the same buffer.

Molecular markers used for the calibration were thyroglobulin kDabovine IgG kDaovalbumin 44 kDamyoglobin 17 kDa and vitamin B12 1. Protein content was quantified by means of the Bradford method [30] using bovine serum albumin for the standard curve.

Plant Physiol and Biochem. Abstract Two phosphatase isoforms from roots of the common bean Phaseolus vulgaris L. Introduction Phosphorus P is one of the major elements found in plants.

Results and Discussion The common bean, like other plant species, responds to Pi starvation with an increase in the activity of acid phosphatases in roots and leaves [6, 20, 21].

Meaning of “preparativa” in the Spanish dictionary

Experimental section Plant materials Phaseolus vulgaris L. Determination of protein Protein content was quantified by means of the Bradford method [30] using bovine serum albumin for the standard curve. Barranca del Muerto No.