摘要:

Polyethylene glycol (PEG) was substituted for ammonium sulfate in the gradient solubilization chromatographic procedure for the purification of proteins. Preliminary studies utilizing this method for fractionation of a crude lysyl hydroxylase preparation yielded an increase both in purity and in recovery of activity when compared with fractional precipitation with PEG. Turbidity, evident in resuspended fractional precipitates, was not found in PEG gradient solubilization fractions containing enzymatic activity.

ISSN:0032-7484    

DOI:10.1080/00327488008061714

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

These studie s were done to determine four basic intrinsic properties of poly(U)-agarose affinity columns. Specificity of binding studies demonstrated that binding to these columns is highly specific with >90% complementary binding and ⩽3% noncomplementary binding. Sensitivity of binding studies indicated that a minimum sequence of 10 adenylates is required for detectable complementary binding. Selectivity of binding studies revealed that nonsequential adenylates in native RNAs and randomly distribut edadenylates in synthetic poly(A)-poly(C) co-polymers did not bind to poly(U)-agarose affinity columns. Whereas, affinity of binding studies demonstrated that A=U complementary base pairing is independent of chain-lengths of ⩾25 a denylates and dependent of chain-lengths of <25 adenylates. Thus the data demonstrates that poly(U)-agarose affinity chromatography is scientifically sound and expedient for thedetection and isolation of poly(A)-containing cellular and viral RNAs.

ISSN:0032-7484    

DOI:10.1080/00327488008061715

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

A simple chemical procedure for the preparation of four common ribonucleoside 5-λ-[32p] triphosphates of high specific activity (up to 10 Ci/mmole) based on the condensation of orthophosphoric acid with the corresponding nucleoside 5-diphosphate in the presence of ethyl chloro-formate as well as the methods of purification and identification of the products are described.

ISSN:0032-7484    

DOI:10.1080/00327488008061716

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

A convenient synthesis of erucyl CoA from N-hydroxysuccinimide ester of erucic acid and coenzyme A is reported. The product has been fully characterized from its spectral and hydrolysis data. The yield is superior to the acid chloride or mixed anhydride methods where side reactions seem to reduce the yield.

ISSN:0032-7484    

DOI:10.1080/00327488008061717

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

A rapid and simple procedure for the preparation, directly from clotted whole blood, of bovine fibrin free of cell membrane contaminants is described. The procedure exploits the ability of detergents to solubilize and extract cell membrane proteins from homogenized clot fragments. The non-denaturing detergents, TX-100* and DOC, used in sequence, or the denaturing detergent, SDS, used alone, proved equally effective. The resulting fibrin has the solubility properties and the subunit composition on SDS gels of highly cross-linked fibrin.

ISSN:0032-7484    

DOI:10.1080/00327488008061718

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

The most widely used methods for preparing erythrocyte membranes are time-consuming, since they involve repeated steps of centrifugation and washing. We have found that membranes can be prepared in a rapid and simple way by molecular-sieve chromatography on a Sepharose® 4B column. Two sizes of columns have been used, one 9-ml column to prepare about 1 mg of membrane proteins in 10 min and one 2-litre column for larger amounts (up to 200 mg in 15 min). The optimal pH and phosphate concentration for the separation of hemoglobin from the membranes were found to he 7.5 and 5 mM, respectively. Sodium dodecyl sulphate (SDS) electrophoretic analysis of the membrane proteins prepared by centrifugation and by this new method shows similar protein compositions.

ISSN:0032-7484    

DOI:10.1080/00327488008061719

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

A gentle method for concentrating very dilute protein solutions is described. The high capacity of aminohexylagarose in adsorbing different proteins is utilized to handle small or large amounts of protein with practically no losses in material or activity. To concentrate very dilute protein solutions as they occur during purification procedures e. g. of enzymes, a one-step non-inactivating nethod is needed that may easily be integrated into the purification programm.

ISSN:0032-7484    

DOI:10.1080/00327488008061720

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

A cyanogen bromide digest of troponin C from rabbit skeletal mus-cle yields two biologically active Ca2+binding peptides. A simple, one-step method for the simultaneous purification, in high yield, of these two peptides is presented.

ISSN:0032-7484    

DOI:10.1080/00327488008061721

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

摘要:

L-[1-14C]lysine was prepared using a technique based on the differential solubility of the diastereomeric salts of DL-lysine with L-glutamic acid. D-[l-14C]llysine was prepared by the enzymatic removal of residual L-[l-14cl]ysine from the reaction mixture remaining after the preparation of L-[1-141- C]lysine. The products were tested with L-lysine decarboxylase and by column chromatography. L-[l-14C]lysine was obtained in 60% yield with a steroisomeric purity of 95.1%, and D-[l-14cllysine was obtained in a slightly higher yield with a steroisomeric purity of 97.5%.

ISSN:0032-7484    

DOI:10.1080/00327488008061722

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor

ISSN:0032-7484    

DOI:10.1080/00327488008061723

出版商:Taylor & Francis Group    

年代:1980

数据来源: Taylor