摘要:

Multiple techniques for solid phase adsorption of 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) were evaluated. Both the porous polystyrene divinylbenzene matrices (BioBeads SMTM) and Extracti GelTMD reduced CHAPS to significantly below its critical micellar concentration while Extracti-GelTMremoved CHAPS to below detectable limits. Bio-Bead extraction of CHAPS correlated with the surface area of the bead type. SM-16 beads, with the largest effective surface area, removed nearly 97% of the detergent. For a given amount of detergent and mass of Bio-Beads, the ratio of sample to total bead volume significantly affected CHAPS adsorption. Total protein recovery with the Extracti-GelTMwas approximately 97%. Protein recovery in the samples treated with Bio-Beads varied from 56-95 %. Chromatographic rather than batch processing yielded optimum recoveries. CHAPS can be effectively removed from dilute protein solutions by solid phase adsorption and this technique offers significant advantages over standard dialysis or gel filtration methods.

ISSN:0032-7484    

DOI:10.1080/10826068908544892

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

摘要:

The mitochondrial hexokinase from rat brain, selectively released from mitochondria by the action of glucose 6-phosphate, can be purified to greater than 90% homogeneity by a single affinity chromatography step on Affi-Gel Blue; the Cibacron Blue F3GA ligand bound to this matrix serves as an analog of ATP, the normal substrate for the enzyme, and selective elution is accomplished using glucose 6-phosphate which is a competitive ligand vs. ATP. With this and other modifications to the previously described procedure (Ref. 8), highly purified enzyme is readily obtained in good yield and with retention of the ability to rebind to mitochondria.

ISSN:0032-7484    

DOI:10.1080/10826068908544893

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

摘要:

Zetaprep mass ion-exchange media represent a rapid and efficient chromatographic tool in the separation of proteins, in place of the conventional agarose or cellulose-based gels. We adopted this method, combined with classical steps, to purify to homogeneity human recombinant interleukin 1β (IL-1β) produced from E. Coli and from S. cerevisiae. An anion exchanger QAE-ZetaPrep was used to achieve a rapid partial purification of both proteins. The IL-1β purification was completed by gel permeation chromatography on Sephadex G-50. When the protein was produced from yeast, an intermediate chromatographic step on a hydroxylapatite column was also necessary.

ISSN:0032-7484    

DOI:10.1080/10826068908544894

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

摘要:

We have developed a procedure for the simultaneous purification of DNA topoisomerase I and II from calf thymus. Both enzymes were first extracted from isolated nucleoprotein complexes. After batchwise chromatography on hydroxylapatite the two enzyme activities were separated on a FPLC phenylsuperose column. The enzymes were further purified by a second chromatography on phenylsepharose (topo I) or FPLC Mono Q (topo II). The purification can be finished within three days, yielding 0.5-1.0 mg quantities of homogeneous, enzymatic active preparations of the two proteins from 200 g of starting material.

ISSN:0032-7484    

DOI:10.1080/10826068908544895

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

摘要:

A method is described to purify pancreatic carboxypeptidases B (CPB), removing contaminating endoproteinases that interfere with use of CPB for carboxy-terminal analysis or modification of proteins. The separation uses zinc chelate chromatography and is based on the property that CPB has higher affinity for immobilized zinc ions than do serine proteinases such as trypsin and chymotrypsin. which are abundant endoproteolytic activities in pancreas. CPB preparations are loaded onto a column of iminodiacetic acid-Sepharose that has been saturated with ZnCl2. A step gradient with buffers of decreasing pH is used to elute bound proteins. CPB elutes at a lower pH than do the serine proteinases.

ISSN:0032-7484    

DOI:10.1080/10826068908544896

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

摘要:

A method is proposed for the isolation (and purification) of enzymes, with retention of their activity, from solutions or gels of preparative PAGE runs. It is based on the inclusion of Sephadex G-25 as a supporting medium for a collector buffer in otherwise normal disc-PAGE gels. The collector buffer has a lower pH and higher concentration than the stacking gel buffer. This makes the proteins concentrate in a very narrow, slowly moving band in the Sephadex on electrophoresis, and makes their recovery easy. The method is illustrated by the isolation of aldehyde oxidase from potato extracts (which was unsuccessful by classical methods), and of one isoenzyme from commercial lipoxygenase after preparative PAGE. Recovery of chicken egg albumin after PAGE was over 90%.

ISSN:0032-7484    

DOI:10.1080/10826068908544897

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

摘要:

Polymer 3520, a non-polar styrene divinylbenzene polymer, provides a simple way to purify calmodulin (CAM) from soybeans. This polymer, which selectively adsorbs CAM by hydrophobic interaction within the polymer matrix, contains no exchangeable groups; thus, interaction with CAM requires no Ca++ions, and elution is achieved with 50% ethanol. Purification by this form of reversed-phase liquid chromatography is a substantial improvement over the conventional method, which requires high salt in elution buffers. CAM in soybean meal is first extracted with 80% ethanol in the presence of EGTA at room temperature and then chromatographed directly on a polymer 3520 column to yield pure CAM. Addition of non-ionic detergent (Nonidet P-40) to the ethanolic extract helps to separate extraneous proteins, lipids, sugars, and isoflavones. Such isolated CAM migrates as a single band during polyacrylamide gel electrophoresis and reversed-phase HPLC, and it retains activity stimulatory to phosphodiesterase.

ISSN:0032-7484    

DOI:10.1080/10826068908544898

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

ISSN:0032-7484    

DOI:10.1080/10826068908544899

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor

ISSN:0032-7484    

DOI:10.1080/10826068908544900

出版商:Taylor & Francis Group    

年代:1989

数据来源: Taylor