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PubMed1998

High-resolution polyacrylamide gel electrophoresis of carbohydrates derivatized with a visible dye.

A technique for carbohydrate analysis that is both inexpensive and easily performed is currently unavailable. In this communication we address the problem and have outlined a method for labeling saccharides with a visible dye, 4-amino-1,1'-azobenzene-3, 4'-disulfonic acid, which has an absorption maximum of 489 nm and an extinction coefficient of 37,615, to facilitate visible detection at low levels. The visible dye was coupled by a reductive amination to different sugars. The sugar-dye adducts were then separated by electrophoresis on alkaline polyacrylamide gels. The gels were scanned with a densitometer, and visible sugar-dye adducts were qualitatively analyzed by identifying them according to their mobilities. The sugar-dye adducts were quantified by determining their densitometric volume. The kinetics of the reductive amination reactions, performed at 37 degrees C, were different for each of three saccharides tested. The rate constants for glucose and fucose were 1.31 times greater and 1.8 times greater, respectively, than that of maltotriose. The reductive amination reactions were essentially complete after approximately 16 h under the given experimental conditions. A linear dose-response relationship was observed between the amount of sugar (monosaccharide, trisaccharide, or heptasaccharide) in the reductive amination reaction. The quantity of saccharide-dye adduct that could be visually detected for glucose, maltotriose, and maltoheptaose, was 25, 25, and 50 nmol, respectively. Sugar-dye adducts were separated from one another by varying the acrylamide concentration in the polyacrylamide gels. Sugar-dye adducts of monosaccharides, disaccharides, trisaccharides, and heptasaccharides were separated on alkaline 30% polyacrylamide gels with mobilities of 0.778, 0.667, 0.639, and 0.375. Adducts of glucose, fucose, galactose, and mannose were separated with mobilities of 0.844, 0.833, 0.820, and 0.810, respectively, on a 30 to 40% gradient polyacrylamide gel. Adducts of glucose and glucose derivatives were separated on a 35% polyacrylamide gel. This technique provides an inexpensive and easily performed method of carbohydrate analysis to laboratories that do not have the highly trained personnel nor the expensive equipment needed for other methods of carbohydrate analysis. The method is most applicable to research problems where sensitivity (20 pmol) is not a problem. The simplicity of the method also makes it easily incorporated into teaching laboratories.

Carbohydrates↗
PubMed1998

Kinetic modeling for macromolecule loading into crosslinked polyacrylamide hydrogel matrix by swelling.

A kinetic model was proposed to characterize the swelling phenomenon of polyacrylamide hydrogel and to quantify and predict the loading of insulin into the hydrogel by swelling. Polyacrylamide hydrogel and porcine insulin were used in the study. During swelling, the insulin concentration in the hydrogel was found to be higher than that in the loading solution, which could be attributed to ionization of the ionic networks, Donnan exclusion, and the possible ionic interactions between the anionic carboxylic pendants and cationic insulin. The experimental results demonstrated that the proposed kinetic model was able to describe the swelling kinetics of polyacrylamide hydrogel and the loading kinetics of insulin by using only two constants [input rate (Kin) and output rate (Kout)]. The experimental values of Kin and Kout were found to highly depend on the concentration of HCl. As medium pH declined (because of the addition of HCl), the degree of swelling decreased and the insulin loading amount in the hydrogel was reduced. A linear log-log function was observed between Kin and the volume fraction of HCl. The Kout values also decreased with the addition of HCl, but remained constant after more than 1% (v/v) of HCl (0.01 N) was added. The proposed model was able to characterize the swelling kinetics of polyacrylamide and predict the loading dose of insulin in the polyacrylamide hydrogel by swelling.

Acrylic Resins↗
PubMedDate not supplied

Severe granulomatous inflammatory response induced by injection of polyacrylamide gel into the facial tissue.

Facial soft tissue augmentation has been performed using different materials. We describe a woman who received polyacrylamide gel injections for cosmetic reasons and developed a severe granulomatous inflammation paranasally. Because the patient did not mention the cosmetic intervention, the diagnosis of polyacrylamide gel-induced granuloma was complicated. The distinctive histopathological findings led to the correct diagnosis despite sparse clinical information. Since complete surgical excision was not feasible, she was treated with repeated multiple local injections of triamcinolone acetonide. Polyacrylamide gel may have favorable properties for facial tissue augmentation, but a severe granulomatous inflammatory response induced by injection of polyacrylamide gel may occur. Before treatment with polyacrylamide gel injection this complication should be disclosed to the patient.

Acrylic Resins↗
PubMed1997

Capillary electrophoresis of DNA fragments in 9 to 20% uncrosslinked polyacrylamide gels: unique separating capacity hypothetically related to maintenance of random-coil DNA conformation independently of gel concentration.

DNA fragments (0.1 to 2 kb) were separated by capillary electrophoresis (CE) in 9 to 20% uncrosslinked polyacrylamide gels with a resolving power ranging from 3 to o.1 million theoretical plates/meter across that DNA size range. The unique feature of electrophoresis in 18 to 20% uncrosslinked polyacrylamide is that it provides a method capable of resolving charge isomeric species of DNA fragments (0.4 to 2 kb), confirming a previous report by Heiger et al. [Heiger DN, Cohen AS, Karger BL. J Chromatogr 516 (1990) 33-48]. A similarly unique resolving capacity of uncrosslinked polyacrylamide gels for DNA previously reported is that for heteroduplex DNA [Pulyaeva H, Zakharov SF, Garner MM, Chrambach A. Electrophoresis 15 (1994) 1095-1100] matched by crosslinked gels only in the presence of denaturants [Peeters AV, Kotze MJ. PCR Methods Appl 4 (1994) 188-190; Ganguly A, Rock MJ, Prockop DJ. Proc Natl Acad Sci USA 90 (1993) 10 325-10 329]. A clue as to the cause of that unique resolving capacity of crosslinked polyacrylamide is provided by the finding in the present study of a single, gel concentration independent KR [retardation coefficient, d(log mobility)/d(gel concentration)] for the DNA fragments, which contrasts with the decrease of KR with gel concentration observed for crosslinked polyacrylamide across a wide concentration range [Orban L, Chrambach A. Electrophoresis 12 (1991) 241-246; Tietz D, Chrambach A. Electrophoresis 14 (1993) 185-190]. Since the decrease of KR with gel concentration correlates with a decrease in equivalent molecular radius [Tietz D, Chrambach A. Electrophoresis 14 (1993) 185-190], it has been interpreted as being due to the transition from a random-coiled to a stretched DNA conformation upon passage through gels of increasing concentration. Since in uncrosslinked gels the decrease of KR does not occur, it is correspondingly assumed that the random-coil conformation of DNA is maintained in those gels in the investigated concentration range up to 20%. The maintenance of random-coil conformation [Tietz D, Chrambach A. Electrophoresis 14 (1993) 185-190]. The effect of denaturants in allowing for resolution of heteroduplex DNA in crosslinked gels [Peeters AV, Kotze MJ. PCR Methods Appl 4 (1994) 188-190; Ganguly A, Rock MJ, Prockop DJ, Proc Natl Acad Sci USA 90 (1993) 10 325-10 329] supports that hypothesis of the enhanced resolving power of electrophoresis in gels that maintain random-coiled DNA within the gel concentration range used.

Acrylic Resins↗
PubMed2006

Efficacy and safety of polyacrylamide hydrogel for facial soft-tissue augmentation in a 2-year follow-up: a prospective multicenter study for evaluation of safety and aesthetic results in 101 patients.

BACKGROUND: Polyacrylamide (Aquamid) is a nonabsorbable soft-tissue filler. It consists of a polyacrylamide hydrogel. Safety and aesthetic results after injection of polyacrylamide hydrogel for facial soft-tissue enhancement were analyzed in a clinical trial. METHODS: In a prospective multicenter study, 251 patients were enrolled for injection of polyacrylamide hydrogel (Aquamid). The results of the first year follow-up have been described previously. One hundred one of 251 initially enrolled patients could be recruited for follow-up 24 months after the first injection. Standardized photographs were taken, and blood and urine samples were collected before treatment and during follow-up visits. Common sites of injection were nasolabial folds, lips, glabella folds, and other sites. The amount of injected gel ranged from 0.2 to 12 ml. RESULTS: A course of 24 months was followed in 101 patients. Results were judged to be good or very good by the investigators in 93 patients. Eighty-six patients judged the aesthetic outcome as satisfactory or very satisfactory. No severe side effects were observed during the 2-year follow-up period. Transient local reactions that resolved spontaneously were detected in several patients. In one case, burning sensations after lip augmentation were reported lasting up to the 24-month follow-up visit. CONCLUSIONS: Polyacrylamide hydrogel (Aquamid) yielded satisfying aesthetic results in more than 90 percent of patients. There was no difference in efficacy between 12 and 24 months' follow-up. No adverse soft-tissue reaction was observed. The study group may be biased, as patients with bad results may have refrained from further follow-up. Long-term follow-up is necessary.

Acrylic Resins↗
PubMed1979

Acrylic microspheres in vivo. I. Distribution and elimination of polyacrylamide microparticles after intravenous and intraperitoneal injection in mouse and rat.

Microspheres of 14C-labeled highly cross-linked polyacrylamide (mean diameter 0.25-0.30 micron) have been prepared by emulsion polymerization. The label was introduced in the polymer via the cross-linking monomer. N,N'-[14C]methylenebisacrylamide. The microparticles were used to follow qualitatively and quantitatively the distribution and the fate of polyacrylamide in mouse and rat after i.v. and i.p. injection. The polyacrylamide particles are rapidly cleared from the circulation (t 1/2 in rat approximately 40 min) by macrophages of the reticuloendothelial system. They are mainly (about 80%) found in the liver and spleen both after i.v. and i.p. injection (4.1 mg given totally to mice weighing 20-25 g). They can also be detected early (1 hr after i.v. injection) in the bone marrow, and particle aggregates are also initially found in the lungs, although no respiratory problems were noted. After about 16 weeks, the radioactivity rapidly decreases in the liver and spleen, with t 1/2 10 to 14 weeks and 15 to 24 weeks, respectively, depending on the route of administration. Radioactivity in the gut and gut walls detected 2 months after injection suggests that the polyacrylamide is slowly metabolized. No toxic effects-apart from transient hepatospleenomegaly-were detected in mice 45 weeks after they were given a total of 9.8 mg of polyacrylamide.

Acrylamides↗
PubMed1978

Adhesion of chicken hepatocytes to polyacrylamide gels derivatized with N-acetylglucosamine.

Complex carbohydrates on the surfaces of eukaryotic cells are thought to participate in a wide variety of cell-cell interactions. A model system has therefore been developed to study these processes. In the present experiments, the ability of chicken hepatocytes to recognize and adhere to sugars covalently linked to polyacrylamide gels was investigated. The gels were snythesized by two methods. Type I gels were prepared from a co-polymer of an active ester of acrylic acid (N-succinimidyl acrylate), acrylamide, and bisacrylamide. The "activated" polyacrylamide gel was then treated with the desired ligand containing an amino group, such as 6-aminohexyl O- or S-glycoside. Type II gels were formed by treating similar ligands with acryloyl chloride, followed by co-polymerization of the resulting N-substituted acrylamide with acrylamide and N,N'-methylenebisacrylamide. These polyacrylamide derivatives offer many advantages for studies with intact cells. They are not toxic to any cell type studied, can be cast in any desired shape, are transparent and stable over a wide range of pH values, and contain no cationic and low to negligible levels of anionic charge (charged groups can be introduced if desired), and the polyacrylamide matrix is stable to common biological agents such as bacteria and enzymes. In addition, type I gels can be synthesized using a broad range of molecules containing amino groups, such as glycopeptides, proteins, etc. The hepatocytes were prepared by collagenase perfusion of intact chicken livers. The rate and extent of adhesion of the cells to the derivatized gels was determined by measuring lactate dehydrogenase in these cells. This enzyme was also used to assay viability and cell "leakiness." At 37 degrees C, 70 to 100% of the cells adhered within 60 min to gels derivatized with N-acetylglucosamine, i.e. gels derivatized with 6-aminohexyl 2-acetamido-2-deoxy-beta-D-glucopyranoside (or the corresponding thioglycoside). By contrast, less than 5% of the cells adhered to polyacrylamide or to gels derivatized with 6-aminohexanol or the 6-aminohexyl glycosides of beta-D-glucose, beta-D-galactose, alpha-D-mannose, beta-D-maltose, beta-D-melibiose, beta-D-cellobiose, and (alpha or beta)-D-lactose. Kinetic studies with the chicken hepatocytes and N-acetylglucosamine gels showed that cell-gel binding was dependent upon Ca2+ and was decreased at low temperatures. Binding was inhibited by N-acetylglucosamine or by glycosides of this sugar, the most effective inhibitor being orosomucoid (alpha1-acid glycoprotein) pretreated with sialidase and beta-galactosidase. The cell surface receptor(s) involved in this interaction is not known, but may be related or identical to the chicken liver binding protein described by Lunney and Ashwell (Lunney, J., and Ashwell, G. (1976) Proc. Natl. Acad. Sci. U. S. A. 73, 341--343). The present results suggest that this model system should prove useful in delineating cell surface interactions with carbohydrates.

Acetylglucosamine↗
PubMedDate not supplied

Detection of a single base mismatch in double-stranded DNA by electrophoresis on uncrosslinked polyacrylamide gel.

Uncrosslinked polyacrylamide forms gels in the concentration range of 15-40% acrylamide. Electrophoresis in these gels of a commercially available 350 bp heteroduplex DNA preparation separates it from the homoduplex DNA of the same size. The separation is qualitatively equivalent to that previously achieved in a commercial proprietary gel ("Mutation Detection Gel" of AT-Biochem), or in an equivalent 14% T, 0.15% C (N,N'-methylenebisacrylamide) gel, but the mechanical stability of mutation detection electrophoresis (MDE) gels or 0.15% C gels is better than that of uncrosslinked polyacrylamide gels. The separation in any of these three gel media can be carried out in short gel tubes within a few hours of electrophoresis time. In both uncrosslinked polyacrylamide and MDE gel media, the Ferguson plots [log(mobility) vs. gel concentration] and the plots of effective molecular radius vs. gel concentration ("T-plots") of both the heteroduplex and homoduplex DNA indicate an augmented size but similar flexibility upon passage through the gel than exhibited by the components of a DNA standard ladder. Homoduplex and heteroduplex DNA correspondingly exhibit a parallelism of their Ferguson curves in transverse MDE pore gradient gel electrophoresis, suggesting a surface net charge difference, possibly due to a conformational reorientation too subtle to be detected by a shift in the slope of the Ferguson plot, as has been observed once previously with a "kinked" DNA species. The gel fiber radius or length per unit volume of uncrosslinked polyacrylamide and MDE gels do not differ significantly within confidence limits, which are wide compared to unconventionally crosslinked gels, presumably because of their greater swelling.

Acrylic Resins↗
PubMedDate not supplied

The relative separation efficiencies of highly concentrated, uncrosslinked or low-crosslinked polyacrylamide gels compared to conventional gels of moderate concentration and crosslinking.

The joint report [1] has shown that the separation of heteroduplex DNA from homoduplex DNA can be achieved by uncrosslinked polyacrylamide gels or gels of a very low degree of crosslinking (0.15%) with N,N'-methylenebisacrylamide (Bis), while conventional polyacrylamide gels of 2-5% crosslinking with Bis are incapable of such a separation in the absence of added denaturing agents. This result raised the question whether in application to other separation problems the same superiority of uncrosslinked or low-crosslinked polyacrylamide existed. To test that question, Ferguson plots were determined for the members of a DNA ladder (50 to 1000 bp) in polyacrylamide with 0, 0.1, 0.2, 0.3, 0.5% C (Bis), and the separation efficiency function, S, was evaluated in comparison with that in conventional 2-5% C (Bis) gels. S was found to be lower, not higher, in gels of low crosslinking at the respective maximally effective gel concentrations. However, the range of gel concentrations in which gels of low or no crosslinking were effective extended over a range of at least 10% T, while conventionally crosslinked gels were most effective over a range of 3 to 1 units of %T.

Acrylic Resins↗
PubMed1992

Sodium dodecyl sulfate-capillary gel electrophoresis of proteins using non-cross-linked polyacrylamide.

Proteins with relative molecular masses of 14,000 to 205,000 were separated by sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) using non-cross-linked linear polyacrylamide gels on both coated and uncoated fused-silica capillaries. It was determined that viscosity of the acrylamide solution was a major factor affecting column stability with linear acrylamide gels. When the viscosity of the acrylamide solution reaches 100 cP, electro-osmotically driven displacement of the gels is insignificant. Uncoated capillaries provided better resolution, stability, and reproducibility than surface coated capillaries when the concentration of linear polyacrylamide was greater than 4%. At lower gel concentrations, non-cross-linked polyacrylamide is easily displaced from the columns. A calibration plot of log molecular mass vs. mobility with non-linear polyacrylamide was linear, which indicated that resolution was equivalent to that obtained with cross-linked acrylamide. Separations with model proteins indicated that baseline resolution between protein species that vary 10% in molecular mass can be achieved.

Acrylic Resins↗
PubMed2001

Separation of double-stranded DNA fragments by capillary electrophoresis in interpenetrating networks of polyacrylamide and polyvinylpyrrolidone.

Mixtures of two polymers with totally different chemical structures, polyacrylamide and polyvinylpyrrolidone (PVP) have been successfully used for double-stranded DNA separation. By polymerization of acrylamide in a matrix of PVP solution, the incompatibility of these two polymers was suppressed. Laser light scattering (LLS) studies showed that highly entangled interpenetrating networks were formed in the solution. Further systematic investigation showed that double-stranded DNA separation was very good in these interpenetrating networks. With a concentration combination of as low as 2% w/v PVP (weight-average molecular mass Mr = 1 x 10(6) g/mol) + 1% w/v polyacrylamide (Mr = 4 x 10(5) g/mol), the 22 fragments in pBR322/HaeIII DNA, including the doublet of 123/124 bp, have been successfully separated within 6.5 min. Under the same separation conditions, similar resolution could only be achieved by using polyacrylamide (Mr = 4 x 10(5) g/mol) with concentrations higher than 6% w/v and could not be achieved by using only PVP (Mr = 1 x 10(6) g/mol) with a concentration as high as 15% w/v. It is noted that the interpenetrating network formed by 2% PVP and 1% polyacrylamide has a very low viscosity and can dynamically coat the inner wall of a fused-silica capillary. The separation reached an efficiency of more than 10(7) theoretical plate numbers/m and a reproducibility of less than 1% relative standard deviation of migration time in a total of seven runs. The interpenetrating network could stabilize polymer chain entanglements. Consequently, the separation speed was increased while retaining resolution.

Acrylic Resins↗
PubMed1988

Application of Neuhoff's optimized Coomassie brilliant blue G-250/ammonium sulfate/phosphoric acid protein staining to ultrathin polyacrylamide gels on polyester films.

An optimized Coomassie staining procedure, utilizing Coomassie Brilliant Blue G-250 in phosphoric acid/ammonium sulfate, was applied to ultrathin-layer isoelectric focusing in 0.18 mm polyacrylamide gels, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in 0.38 mm polyacrylamide gels, both backed to Gel-Fix polyester supporting films. After isoelectric focusing staining of gelatin and acidic proteins was better with the phosphoric acid/ammonium sulfate procedure than with conventional organic solvent methods. When applied to gels after sodium dodecyl sulfate-polyacrylamide gel electrophoresis the sensitivity of the phosphoric acid/ammonium sulfate method was equal to that on conventional staining but lower than on silver staining.

Acrylic Resins↗
PubMed1991

Information on DNA conformation derived from the Ferguson plot of DNA fragments of up to 9 kb in size, using polyacrylamide gel electrophoresis in a discontinuous buffer system.

The Ferguson plot in polyacrylamide gel electrophoresis (PAGE)(15%CDATD, moving boundary electrophoresis buffer system operative at pH 8.9, 4 degrees C, 8 mA/cm2 of gel) of DNA fragments up to 9.4 kb in size was found to exhibit a linear segment at polyacrylamide concentrations starting at 3% T and undergoing a gradual transition into a concave segment at higher gel concentrations, confirming previous findings by Stellwagen. The larger the DNA, and the higher the gel concentration, the less extended the linear and the more extended the concave segment of the plot. The lowest % T of the linear range for DNA in polyacrylamide remains unknown since mobilities at nongelling concentrations below 3% T have not as yet been measured. As previously suggested, the transition from the linear to the concave segment corresponds to that from the randomly oriented DNA to the anisotropically stretched, "reptating" DNA. For a DNA of 9.4 kb in size, the end of the linear range of the Ferguson plot can be extended from 3.5 to 5% T when 15% DATD rather than 2.5% Bis is used to crosslink the polyacrylamide. Increasing the temperature of PAGE from 4 degrees C to 25 and 50 degrees C widens the linear segment progressively, indicating an increasingly random orientation with rising temperature. When current density is increased from 8 to 40 mA/cm2, the concave curvature of the Ferguson plot of DNA 1 to 9.4 kb in size decreases, suggesting a transition from a "reptating" to a randomly distributed molecule, due to increased Joule heat.(ABSTRACT TRUNCATED AT 250 WORDS)

Buffers↗
PubMed1995

New types of large-pore polyacrylamide-agarose mixed-bed matrices for DNA electrophoresis: pore size estimation from Ferguson plots of DNA fragments.

The average pore size value of gels containing polyacrylamide, covalently linked to agarose, was found to be 30% higher than the value of a regular N,N'-methylenebisacrylamide (Bis) cross-linked gel of the same %T. By increasing the agarose concentration (10% of the total amount of polyacrylamide), gels containing low amounts of acrylamide (1.5-2%) are reproducibly obtained; their pore sizes are 130% larger than the pore sizes of a 4%T, 3.3%C polyacrylamide gel. In general, mixed-bed matrices were found to be more elastic and mechanically stronger than classical polyacrylamide gels since an agarose-induced gelation process takes place during their polymerization.

Acrylic Resins↗
PubMed1995

Detection of giant myofibrillar proteins connectin and nebulin by electrophoresis in 2% polyacrylamide slab gels strengthened with agarose.

We have established an improved method of sodium dodecyl sulfate-polyacrylamide gel electrophoresis to facilitate analysis of giant myofibrillar proteins connectin and nebulin, whose molecular masses are about 3000 and 700 kDa, respectively. This method consists of 2% polyacrylamide slab gels strengthened with agarose and a specific buffer system. Our 2% polyacrylamide slab gels are superior to usual 1.8 or 2% polyacrylamide disc gels in preparation, handling, and resolution.

Animals↗
PubMed1995

Activity staining of mammalian ribonuclease inhibitors after electrophoresis in sealed vertical slab polyacrylamide gels.

A method for detecting the activity of ribonuclease inhibitors (RIs) after nondenaturing polyacrylamide gel electrophoresis and isoelectric focusing was developed. Both types of electrophoresis were performed using vertical slab polyacrylamide gels in the presence of dithiothreitol and in a sealed system. In each system, purified 50 kDa human RI was visualized as a single band by immunoblotting with a specific antibody. RI activity in the polyacrylamide gel slab was detected by sandwiching the gel slab between a cellulose acetate membrane moistened with a solution of bovine pancreatic ribonuclease A and a dried agarose film sheet containing substrate yeast RNA plus ethidium bromide and incubating at 37 degrees C. The ribonuclease penetrated the polyacrylamide gel and digested the substrate RNA in the agarose film. However, if an RI was present in the gel, the enzyme was inactivated by complex formation. Fluorescent bands corresponding to RIs were observed on a dark background under ultraviolet light. This activity staining had a high sensitivity allowing detection of less than 0.6 units of mammalian RIs (corresponding to 5 ng of purified human RI) and produced a sharp band which compared favorably with that obtained on immunoblotting. These electrophoretic techniques appear useful for the investigation of RIs in heterogeneous biological samples.

Animals↗
PubMed1990

Detection of glycoproteins separated by nondenaturing polyacrylamide gel electrophoresis using the periodic acid-Schiff stain.

We observed that published methods for staining glycoproteins in sodium dodecyl sulfate-polyacrylamide gels did not stain glycoproteins separated by nondenaturing polyacrylamide gel electrophoresis. Therefore, the periodic acid-Schiff stain for glycoprotein was adapted for use with proteins analyzed by nondenaturing polyacrylamide gel electrophoresis. Following nondenaturing polyacrylamide gel electrophoresis proteins were denatured in situ by incubation with aqueous 2% sodium dodecyl sulfate, 5% 2-mercaptoethanol, 40% ethanol, and 5% acetic acid at 90 degrees C for 15 min followed by periodic acid-Schiff staining. This modified procedure will detect at least 0.2 micrograms protein-associated carbohydrate. Omission of the periodic acid treatment from the protocol was used as a control to detect nonspecific staining of some proteins. This modified procedure was validated using both purified glycoproteins and extracellular culture fluid containing carbohydrate-associated proteins of Ruminococcus flavefaciens.

Bacteriological Techniques↗
PubMed1975

Isolation of outer membrane proteins of Escherchia coli and their characterization on polyacrylamide gel.

Proteins from the outer membrane of Escherichia coli were studied on a ureadodecyl sulfate polyacrylamide gel by electrophoresis. A polyacrylamide gel containing sodium dodecyl sulfate and urea gave an excellent resolution of outer membrane proteins. Seventeen protein bands were reproducibly observed on a gel. By use of Sephadex G-200, DEAE-cellulose and polyacrylamide gel, eight proteins were purified to near homogeneity. Five of them were found to be heat-modifiable proteins. The behavior of these purified proteins was studied on a polyacrylamide gel under three different electrophoretic conditions, which had been used for the analysis of cell envelope proteins. Thus correspondence was made between these purified proteins and envelope proteins reported by other investigators.

Bacterial Proteins↗