摘要:

AbstractThe study of effects of several parameters on silver staining of proteins has led to the development of a staining method which is simple and reliable, requires only few stable solutions, and can be applied to all gel types such as sodium dodecyl sulfate (SDS) containing polyacrylamide or isoelectric focusing gels. It can be used for ultrathin layers (0.1–0.2 mm) or thicker slab gels (up to 3 mm). With this method not only proteins but also polypeptides of molecular weights as low as 2500 are detectable with high sensitivity. Comparison of isoelectric focusing and SDS‐containing gels and a simple spot test on thin gellayers show that the detection sensitivity depends not so much on the type of proteins but rather on their structure. The redox properties of the gel are important for the staining mechan

ISSN:0173-0835    

DOI:10.1002/elps.1150060302

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractFormulations are presented for 4 to 6 pH unit wide immobilized pH gradients requireing the use of only six available Immobiline buffers, without strong acidic and basic titrants. Recipes are also given for the pH 3–10 rang

ISSN:0173-0835    

DOI:10.1002/elps.1150060303

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractPurified human serum albumin (HSA) from normal adults has been reactedin vitrowith different trioses and hexoses and the reaction products analysed by ultrathinlayer isoelectric focusing (IEF) under native and denaturing (8Murea) conditions. The order of reactivity of these molecules has been found to be: glyceraldehyde>glyceraldehyde 3‐phosphate>glucosamine ≫ glucose ≫ acetaldehyde. By chemical analysis, the number of residues of glyceraldehyde bound to albumin appears high: 130 moles/mole of protein, while the number of exposed lysines is halved:ca.8 in reactedvs.16 in control albumin. It appears that a minimum length for an aldehyde to react with protein amino groups is three carbon atoms. A completely different IEF behavior has been found between “reacted and NaCNBH4reduced”vs.“reacted and rearranged” HSA molecules. The former group, even when extensively reacted with sugars, exhibits a normal surface charge at pH = pI, possibly because, at the acidic pIs of HSAs, the now secondary (sugar‐bound) amino groups are still fully protonated. In the latter group, the bound sugar spontaneously undergoes an Amadori rearrangement followed by secondary reactions, still not fully clarified, which could possibly lead to ring structures (e. g.pyrrolic) with complete loss of positive charge from the sugar‐reacted amino groups. These species, in fact, show a marked

ISSN:0173-0835    

DOI:10.1002/elps.1150060304

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractThe sera of 49 subjects with IgG, IgA or IgM benign monoclonal gammopathy‐components were examined by agarose gel isoelectric focusing followed by Coomassie Brilliant Blue R‐250 staining as well as avidin‐biotin amplified double‐antibody peroxidase labelling after nitrocellulose blotting. IgG‐components gave 2–10 distinct bands within the isoelectric point (pI)‐range of pH 6.5–9.5, IgA‐components were focused into 10–15 bands with pI‐values of pH 4.5–6.5 and the IgM‐components gave 1–2 fractions within the pI‐range of pH 4.5–6.5. The avidin‐biotin peroxidase complex (ABC) technique exhibited some tendency to non‐specific labeling related to the avidin‐biotin component and depending on the relative amounts of the individual proteins. However, the method has a definite potential for the visualization of submicrogram quantities of immunoglobulins blotted to nitrocellulose

ISSN:0173-0835    

DOI:10.1002/elps.1150060305

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractThe isotachophoretic behavior of quaternary benzo/c/phenanthridine, protoberberine and aporphine alkaloids in different electrolyte systems is described. The concentration of the leading ion and the pH value of the leading electrolyte affect the relative effective mobilities of the alkaloids. The system of pH 4.7, containing the leading ion K+(0.005 M) counter ion acetate, and the terminating ion β‐alanine (0.02 M), has been selected for the quantitative determination of the studied alkaloids in model mixtures and plant extrac

ISSN:0173-0835    

DOI:10.1002/elps.1150060306

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractEsterases produced by different bacterial species were separated by conventional electrophoresis (CE) in polyacrylamide agarose gel and by thin‐layer isoelectric focusing (IEF). Although CE revealed the greatest number of esterases and was preferable for their identification by specific hydrolytic activities, the two techniques appeared to be complementary in their resolving power for detection of electrophoretic variants. Consequently, by establishing a direct correspondence between homologous esterase bands resolved by CE and IEF, we have proposed a two‐dimensional electrophoretic profile (2‐DEP) which considerably refined the degree of esterase polymorphism and improved the enzymic differentiation between and within bacterial sp

ISSN:0173-0835    

DOI:10.1002/elps.1150060307

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractA simple method for drying polyacrylamide gels of various size (0.3 mm capillary gels to 3 mm thick slab gels) and composition (T = 5–20 %, C = 0.7–5.3 %, sodium dodecyl sulfate gels, acetic acid/urea gels, isoelectric focusing gels) is described. The gels are destained in the presence of 20 % glycerol, dehydrated in acetone, and dried at room temperature. By this procedure, size reduced, clear or slightly opaque, and flexible sheets are obtained. The dried gels can be used for fluorography, stored for long periods and rehydrated without any cracking, warping, or loss in resolution. The shrinkage of the gels to about 25 % of their original size during acetone‐dehydration results in an increased staining intensity and shorter exposure times for fluorog

ISSN:0173-0835    

DOI:10.1002/elps.1150060308

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractGelatins with various electric charges were added to agarose to control the strength of electroendosmotic flow and its effect on esterase zymograms was studied. Addition of the “basic fraction of gelatin” suppressed electroendosmotic flow. In contrast, addition of the “acidic fraction of gelatin” or “modified gelatin” intensified electroendosmotic flow. Modified gelatin was prepared by acylation of amino groups in the basic fraction of gelatin. The observed effects depended on the amount of gelatin added. The gelatin was shown to control electroendosmosis due to electric charge. In addition, the gelatin was found to exhibit a molecular sieving effect. Electroendosmotic flow and molecular sieving originating from the gelatin resulted in high resolution of the esteras

ISSN:0173-0835    

DOI:10.1002/elps.1150060309

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractRecently, Altland and Altland (Clin. Chem.1984,30, 2098–2103) described three alternative devices for pouring immobilized pH gradients (IPGs) based on simultaneous delivery of a density gradient and a non‐density gradient by using high precision burettes under computer control. One of the expected improvements was an unlimited flexibility for the slope of the non‐density gradient (e. g.the pH gradient) which is stabilized by the density gradient. The human plasma proteins prealbumin and apolipoprotein AI and some of their inherited variants are demonstrated by double one‐dimensional electrophoresis in the sequence polyacrylamide gel electrophoresis followed by isoelectric focusing (IEF) in IPGs of varying slopes in the range of pH 4 to 7. It is shown that the patterns obtained follow the expectations from the gradients drawn on the computer. We conclude that the new devices for pouring IPGs can provide a valuable tool for the easy handling of complex separation problems to be solved

ISSN:0173-0835    

DOI:10.1002/elps.1150060310

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractA method for the determination of amprolium in feed stuff by analytical capillary isotachophoresis with column coupling is described. Up to 20 μl of the raw water extract of amprolium from a feed stuff can be analysed directly without any pretreatment in 15 min. The detection limit reached the level of ppm, the reproducibility of analyses in the followed range of 0.2–1.2 nmoles was 3 rel.

ISSN:0173-0835    

DOI:10.1002/elps.1150060311

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY