Article

[Pattern and nomenclature of serum proteins of the amphibian urodele Ambystoma dumerilii Duges]

May 1978
Comptes rendus des séances de l'Académie des sciences. Série D, Sciences naturelles 286(13):1077-80

Source
PubMed

Authors:

By means of many electrophoresis techniques, the serum of Ambystoma dumerilii was studied. The pattern in cellulose acetate is different from A. mexicanum proteinogram. Direct two-dimensional analysis in acetate/polyacrylamid gel with progressively increasing concentration from 4-30%, gives us the opportunity to observe 41 proteic components. By immunological results, the fastest proteic fraction is equivalent to a globuline with alpha-mobility more than an albumin. A nomenclature for this blood serum is proposed.

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Article

Dry bean cultivar characterisation by isoelectric focusing electrophoresis in polyacrylamide gel

July 2001 · Journal of The Science of Food and Agriculture

A Spanish pinto bean local cultivar ‘Pinta’ was analysed by a standard electrophoretic technique, isoelectric focusing electrophoresis, in order to develop and test a reliable method of solving such food industry problems as fraud, adulteration and the identification and registration of different cultivars of legume seeds. The corresponding dry bean type protein extract was subjected to ... [Show full abstract] isoelectric focusing electrophoresis in polyacrylamide gel (IEF‐PAGE), and the resulting data were then studied with a suitable objective chemometric evaluation technique. Applying the Rasch model to the appropriate electrophoretic results, this Spanish dry bean type was clearly characterised by the order of the chemometric measures attributed by the model to each electrophoretic band in its proteinogram. The same chemometric technique was then applied to three more dry bean cultivars. They also showed their own characteristic order of the electrophoretic bands in their corresponding proteinograms. Consequently, this method could well be a valuable tool for legume seed identification and registration. The problems intrinsic to electrophoresis were solved with the Rasch model techniques. © 2001 Society of Chemical Industry.

Article

Interpretation of capillary zone electrophoresis compared with cellulose acetate and agarose gel ele...

October 2010 · Veterinary Clinical Pathology

Serum protein electrophoresis is widely used for diagnostic and research purposes. Cellulose acetate (CAE) and agarose gel (AGE) electrophoresis are the most frequently used methods, but capillary zone electrophoresis (CZE) is beginning to be used more in veterinary laboratories. However, reference intervals for CZE in animals and comparison studies with the other electrophoretic techniques are ... [Show full abstract] lacking, compromising the diagnostic utility of CZE. The aims of this study were to compare results obtained using CAE, AGE, and CZE; to establish reference intervals for CZE in dogs and cats; and to assess the capacity of CZE to detect abnormalities identified by AGE. Serum samples from 204 dogs, including 104 healthy animals, and 62 cats, including 28 healthy animals, were analyzed using automated systems for CAE, AGE, and CZE. Descriptive statistics and Passing-Bablok and Bland-Altman tests were used to compare results. For each technique, reference intervals were calculated based on results from healthy animals. Concordance between CZE and AGE in detecting pathologic changes was assessed using Cohen's k coefficient. For most protein fractions, values obtained by CAE, AGE, and CZE were significantly different from each other, and constant and proportional errors were often detected. Nevertheless, reference intervals obtained by the 3 techniques overlapped. Moreover, Cohen's k coefficient demonstrated that the capacity of CZE and AGE to detect pathologic changes was comparable. CZE performs comparably to AGE and CAE as long as CZE-specific reference intervals are used for interpretation and distinctive visual patterns for albumin, gaps between fractions, and subpeaks found on CZE tracings are recognized. In addition, CZE offers several technical advantages, such as ease of use and complete automation.

Article

Diagnosis of Pseudo-Arylsulfatase A Deficiency with Electrophoretic Techniques

November 1984 · Pediatric Research

Deficient arylsulfatase A activity in man has long been associated with the neurodegenerative disease, metachromatic leukodystrophy. However, similar deficiency has been noted in clinically normal individuals, and is referred to as the pseudoarylsulfatase A deficiency condition. Although direct quantitative analysis of arylsulfatase A activity failed to differentiate between these two conditions, ... [Show full abstract] analysis of residual arylsulfatase A activity with either Cellogel electrophoresis or isoelectric focusing in polyacrylamide gels now has been shown to distinguish between them unequivocally. With both techniques, cultured fibroblasts from patients with pseudo-arylsulfatase A deficiency showed faint but clear bands of arylsulfatase A activity. Under identical conditions, fibroblasts from patients with metachromatic leukodystrophy showed no trace of activity. These methods can be adapted easily for general laboratory analysis in cases when results from quantitative arylsulfatase A assays are noninformative.

Article

Full-text available

Proteins of linseed (Linum usitatissimum L.), extraction and characterization by electrophoresis

February 1999 · Botanical Bulletin- Academia Sinica Taipei

Reda Sammour

The seed proteins of linseed (Linum usitatissimum L.) were qualitatively and quantitatively analyzed. Qualitative studies were carried out using an array of electrophoretic techniques, including sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), sodium dodecylsulphate porosity gradient polyacrylamide gel electro-phoresis (SDS-Poro-PAGE), two dimension sodium dodecylsulphate ... [Show full abstract] polyacrylamide gel electrophoresis (2-D SDS-PAGE), isoelectric focusing, and mapping gels. Electrophoretic patterns of the proteins extracted with water, buffer, urea, and SDS, and analyzed on SDS-PAGE and Poro-SDS-PAGE under non-reducing conditions, showed six major bands with MWs of 55 kDa, 50 kDa, 47 kDa, 45 kDa, 43 kDa and 41 kDa. After reduction with 2-mercaptoethanol (2-ME), these bands gave rise to large acidic chains with MWs around 40 kDa and small basic chains with MWs around 20 kDa. The pI-values of linseed proteins are distributed over the pH range used, namely pH 310. Mapping gels showed that the major bands were highly heterogeneous. Quantitative estimation of the different protein species in the seed flour indi-cated that the contents of albumin, globulin, prolamin, and glutelin were 197 ± 19, 196 ± 12, 32 ± 7, and 65 ± 6 mg/g seed flour, respectively. Kilodalton (kDa); 2-mercaptoethanol (2-ME); Molecular weights (MWs); Two dimen-sion-PAGE (2-D SDS-PAGE). Abbreviations: SDS-PAGE, sodium dodecylsulphate polyacrylamide gel electrophoresis; SDS-Poro-PAGE, sodium dodecylsulphate porosity gradient polyacrylamide gel electrophoresis; 2-D SDS-PAGE, two dimension sodium dodecylsulphate polyacrylamide gel electrophoresis; 2-ME, 2-mercaptoethanol; PAGIF, polyacrylamide gel isoelectric focusing.

Article

Immunochemical Identification of Normal and Variant Hemoglobins after Electrophoretic Separation and...

February 1984 · Hemoglobin

A highly specific method for the conclusive identification of normal and variant human hemoglobins is described in this communication. The method employs the standard electrophoretic technique in combination with the immunoblot technique using monospecific antisera raised in rabbits. Various hemoglobins such as Hb, S, C, A2, and F were separated on an alkaline polyacrylamide gel or a ... [Show full abstract] cellulose-acetate membrane, and transferred to a nitrocellulose membrane and the individual hemoglobins were identified by the immunoblot procedure. With this method hemoglobins with similar or identical electrophoretic mobilities can be definitively identified with the use of monospecific antisera.

Article

Albumin-bound fluorescence in serum of patients with chronic renal failure

May 1980 · Clinical Chemistry

A band that is strongly fluorescent and migrates electrophoretically with serum albumin is commonly found in electrophoretograms of sera from patients with chronic renal failure. We sought to determine whether the fluorescence originates from binding of certain still-unidentified metabolites or drugs, from an abnormal albumin species, or from some other protein entity. Molecular-exclusion column ... [Show full abstract] chromatography, polyacrylamide gel isoelectric focusing, and cellulose acetate electrophoresis, along with results of charcoal treatment and alcohol extraction, provided evidence that the fluorescence comes from fluorescent ligands tightly bound to albumin. The fluorescent intensity of the albumin fraction, isolated by molecular-exclusion chromatography, coincides with the albumin-associated fluorescence determined electrophoretically and with the intensities of the fluorescence emission spectrum for serum. A fluorescent species with an emission maximum of 415 +/- 5 nm, separated by thin-layer chromatography, appears to account for the increased serum fluorescence.

Article

Protein Concentration in Interstitial and Lymphatic Fluids from the Subcutaneous Tissue

February 1977 · Acta Physiologica Scandinavica

The protein content of interstitial fluid and lymph from the same tissue was determined fluid samples of nanolitre volumes being taken from the subcutaneous tissue of rabbits by micropuncture. In the same area lymph was collected from lymphatic vessels of 30-100 mum. An electrophoretic technique in polyacrylamide gels in glass capillaries was used for the analysis of proteins. The gels were ... [Show full abstract] stained and then scanned on a microscope equipped with a moving table and photomultiplier. The area under each protein band was calculated from the recorded densitograms. The method required only a few nanolitres of fluid for analysis. Using solutions of known protein concentrations the relationship between the amount of protein and the area under the densitogram band was investigated. This relationship was found to be linear, making it possible to quantify the protein content of both interstitial fluid and lymph samples. The interstitial fluid/plasma concentration ratios for albumin, transferrin, globulins and total proteins were 0.42, 0.42, 0.32 and 0.37. The albumin/globulin ratio in interstitial fluid was found to be 1.60 of the plasma value although the values obtained for lymph were similar to those found for interstitial fluid. The similarity between the protein concentration of interstitial fluid and lymph indicated that the endothelial lining of the terminal lymphatics did not restrict the passage of macromolecules into the lymphatics.

Article

Principles of isotachophoresis

January 1982 · Fresenius Journal of Analytical Chemistry

Although the theoretical groundwork for isotachophoresis was laid down as long ago as 1897, it is only really in recent years that this technique has been exploited for analytical or preparative purposes. Isotachophoresis is not only the newest, but also generally speaking the least well understood of the modern electrophoretic methods. In the past 10 years, several articles have appeared which ... [Show full abstract] describe the principles and theory of isotachophoresis. Notable are the works of the Everaerts group in Eindhoven, who brought about the practical realisation of isotachophoresis as an analytical procedure in capillary tubes in the late 1960's. However, the mathematics involved in the theory of isotachophoresis may be off-putting to many scientists of other disciplines who are attempting to gain a basic understanding of the principles. Many shorter reviews by other authors during the past decade have not provided sufficient information for a clear picture of the principles. In the present article, we have attempted to find a suitable compromise between extensive and detailed, but too theoretical treatments, and brief description which leave too much unsaid. It is true to say that isotachophoresis has become associated with the technique in capillary tubes, and in our discussion of instruments, we restrict ourselves to such a system. However, in common with all other electrophoretic techniques, isotachophoresis can be performed in a supporting phase, such as polyacrylamide or agarose gel, whereby these procedures have more applicability from the preparative rather than from the analytical point-of-view. The literature list at the end of this article provides some key references for those interested in following-up particular aspects of isotachophoresis, ranging from a more detailed theoretical background, to applications in various fields. It is perhaps useful to provide a short definition of isotachophoresis at this point. An isotachophoretic migration is one in which a mixture of ions of similar charge quality separate into discrete zones of regulated concentration behind an ionic zone of higher mobility than all ions in the mixture (leading ion) and in front of an ion of lower effective mobility than all ions in the mixture (trailing or terminating ion). In the pseudo-steady state, all zone boundaries migrate with identical velocity through the electric field. A characteristic of isotachophoretic separations is the fact that a common buffering counter ion is employed, which defines the pH and hence the effective mobility and migrating order of the zones in the steady state stack. Those conversant with electrophoretic methods will recognise in this discontinuous electrolyte system the concentrating stage of the disc electrophoresis of Ornstein and Davis. This procedure is in fact isotachophoresis. In disc electrophoresis, however, the actual analysis is derived from the second stage, which is zone electrophoresis. Isotachophoresis is used here simply to concentrate the protein zones to improve the resolution of the zone electrophoretic separation. Finally, it is worth pointing out that isotachophoresis is known by several names, which provides a certain degree of confusion for literature searches. Examples are displacement electrophoresis, ionic migration technique, cons electrophoresis, omegaphoresis, transphoresis, steady-state stacking. Although articles still appear using these nomenclatures, the name isotachophoresis, derived from the equal migrating velocity of all zones in the system, is now generally accepted, at least by the major contributors to research in this field.

Article

Human serum albumin variants: determination and repartition of allotypes in 24 cases of bisalbuminem...

October 1976 · Biomedicine / [publiée pour l'A.A.I.C.I.G.]

Weitkamp et al. (1973) using starch gel electrophoresis in three different systems of buffers (pH 5, 5.6 and 6.9) distinguish at least 24 genetically determined variants. In the European area only seven variants have been observed. The study of 24 cases of bisalbuminemia observed in the french population by acetate electrophoresis at pH 8.6 and polyacrylamide agarose gel electrophoresis at pH 8.7 ... [Show full abstract] and pH 6.9 can distinguish three slower variants than the normal albumin. The most frequent variant is the B type observed in 16 cases. The other types are the Pollibauer type observed in 7 cases, and the Gainsville type in one case only. The incidence of these variants can be estimated to 0.7 p. 1000 individuals.

Article

Increased globulin fractionation by an improved vertical gel electrophoretic technique

August 1964 · Journal of Laboratory and Clinical Medicine

A LORBER

An apparatus and technique for vertical gel electrophoresis have been developed which have achieved excellent fractionation of serum proteins. In some pathologic specimens, more than 27 distinct fractions have been distinguished. The acrylamide gel "cartridge" is premolded independent of the apparatus. This eliminates needless tie-up of the apparatus during the gelling procedure and permits ... [Show full abstract] defective gels to be discarded prior to electrophoresis. In addition, gel "cartridges" in a variety of pore sizes may be conveniently stored for use with proteins of varying size. The use of thicker gels enhances columnation of the protein fractions so that even trace amounts of protein are more readily apparent. The large, premolded sample troughs also facilitate introduction of the protein specimen into the gel. Serum specimens of several connective tissue disorders were studied by the technique described, and striking differences in the electrophoresis patterns were demonstrated. Distinctions were noted primarily in the gamma globulin zone. Normal serum specimens, in contrast to pathologic specimens, did not yield a large number of distinct fractions. Protein interaction with DNA was readily demonstrated by this electrophoretic technique. On comparing two identical serum specimens, one of which was preincubated with DNA, considerable loss of one of the protein bands in the gamma globulin zone was demonstrated in the DNA treated specimen. The protein band corresponding to the DNA interacting fraction could also be dissociated in the gel by treatment with low molecular weight thiol compounds. This electrophoretic method can thus be used to identify and quantitate protein reactions with DNA and other compounds.

Article

Early synthesis of uterine proteins after a decidual stimulus in the pseudopregnant rat

September 1976 · Biology of Reproduction

The synthesis of soluble proteins in the uterus of the pseudopregnant rat after decidual induction has been examined by means of a polyacrylamide electrophoretic technique with double labeling incorporation of amino acids. It has been shown by the incorporation of [3H]leucine into soluble proteins that the endometrial tissue synthesizes a single protein band 1 h after the decidual stimulus. This ... [Show full abstract] protein migrates ahead of the bulk of the proteins, and slightly faster than serum albumin during the 6 per cent polyacrylamide electrophoretic run. The protein(s) neither shows up when the whole uterus is examined 1, 2, 3, 4 and 5 h after decidual induction nor in the endometrium on day L3 of pseudopregnancy, the day on which the uterus is insensitive to the decidual stimulus. It is suggested that this protein is synthesized only when the endometrial cells differentiate to form the decidual tissue. The antimesometrial region of the endometrium previously subjected to the decidual stimulus seems to show a higher capacity to synthesize this protein than in the mesometrial region of the same stimulated tissue.

Article

Use of pore size concentration gradient in polyacrylamide electrophoresis

June 1968 · Clinica Chimica Acta

A technic has been devised for the preparation of a pore size-concentration gradient at the sample end of polyacrylamide gel in the disc process. It enables one to achieve added resolution of the globulins at the slow end of the electrophoretic system. What appears to be two high concentration proteins in the more conventional procedures separates to yield several diverse proteins stemming ... [Show full abstract] primarily from the last band. The injection procedure which dilutes the top of the gel to achieve the gradient represents less effort in achieving better resolution than the system from which it was modified.