Cellulose acetate membrane electrophoresis

Polyacrylamide gel electrophoresis (PAGE) and cellulose acetate membrane electrophoresis (CAME) were applied to distinguish escolar and oilfish from 27 commercial fish based on muscle protein differences (Ochiai et al., 1984). Myogen fractions from the muscles were subjected... 

When analyzing the remains of the cooked fish (identified as Yongeichthys nebulosus), TTX was demonstrated by TLC, high-performance liquid chromatography (HPLC), and cellulose acetate membrane electrophoresis. Toxicity was assayed by using ICR (Institute of Cancer Research) strain adult male mice and the toxicity score was 25 MU/g in fish muscle. The synergistic effect of uremia and TTX is obvious in this incident in which the patient and her... 

Storage of human saliva, serum, and duodenal secretion transformed the amylase fractions on cellulose acetate membrane electrophoresis into more-anionic forms. Incubation with lectins, proteases, D-glucosidases, neuraminidase, and some effectors did not modify this conversion, which was promoted by rising temperature and pH values. Increasing concentrations of ammonium ions delayed the transformation of amylolytic fractions, thus indicating non-enzymatic deamidation as the reason for amylase isoenzyme development. A change of molecular weight could be excluded. 

Recently, microchip electrophoresis was applied to GAG analysis using ethidium bromide as a fluorescent dye. In particular, separation times were reduced to 150 s, while sensitivity remained comparable to that of conventional electrophoretic methods that rely on cellulose acetate membranes [47]. 

The high sensitivity of ETA—AAS for Cu has stimulated the development of methods to measure concentrations of the Cu carrier species in biological fluids. Delves [7] analysed the Cu content of the protein fractions separated from 2 pi volumes of serum by cellulose acetate membrane (CAM) electrophoresis. The separated protein bands were cut from the CAM and placed directly into the ETA via a 6 mm x 1 mm hole cut in the wall of the graphite tube. Calibration was achieved by adding 2 pi volumes of aqueous standards to 8 mm x 6 mm strips of CAM. Background correction was essential. Approximately 94% of the Cu was located in the a2 band, where carulo-plasmin would run, whereas other fractions contained less than 5% of the total serum Cu. The recovery of Cu after electrophoresis was quantitative, 99%, and the RSD was 0.086 at 1.74 ng Cu. This method was applied to studies of Cu changes in patients with Menkes Syndrome receiving intramuscular injections of copper as the EDTA complex, and in children with acute lymphoblastic leukaemia. 

Electrophoretic separation on agarose gels or cellulose acetate membranes is the procedure most commonly used to demonstrate LD isoenzymes." After the isoenzymes have been separated by electrophoresis, a reaction mixture is layered over the separation medium. The mixture (typically D, L-lactate> 500mmol/L, and NAD, 13mmol/L, often dissolved in a suitable pH 8.0 buffer) is applied as a liquid or in a gel. The NADH generated over the LD zones is detected either by its fluorescence, when excited by long-wave ultraviolet light (365 nm), or by its reduction of a tetrazolium salt to form a colored formazan. 

Serum LDH isoenzymes can be separated by electrophoresis on agarose gel or cellulose acetate membrane, usually at pH 8.6. After separation, their location is determined by incubation of the support medium in a... 

F23. Friedman, H. 8., A standardized procedure for serum protein electrophoresis on cellulose acetate membrane strips. Clin. Chim. Acta 6, 775-781 (1961). 

A9. Ambler, J., Janik, B., and Walker, G., Measurement of glyc-osylated hemoglobin on cellulose acetate membranes by mobile affinity electrophoresis. Clin. Chem. 29, 340-343 (1983). 

Electrophoresis is a relatively simple and rapid method with high resolution detection of polar compounds like TTX. When 1 jxl of TTX (10 MU, corresponding to 2 jLg) is applied onto a 5 x 18 cm cellulose acetate membrane (Chemetron, Milano), the ion molecules of TTX move toward the cathode with a mobility (Rm) clearly smaller than that of authentic STX. The analysis is performed for 30 minutes in an electrolytic buffer solution of 0.08 mol/liter Tris-HCl (pH 8.7), under the influence of an applied electric field with a constant current of 0.8 mA/cm width. The toxin is visualized in the same manner as described for TLC. 

Cellulose acetate as a medium for electrophoresis was introduced by Kohn in 1958. It was developed from bacteriological cellulose acetate membrane filters and is commercially available as high purity cellulose acetate strips, which are thin and have a uniform micropore structure. 

A simple overlay technique has been reported for the detection of hyaluronidase activity following electrophoresis on cellulose acetate membranes. This sensitive method appears to be specific for e/z fo-jS-D-acetamidodeoxyglucanase activity. 

Interconversions based on a subunit model of the A, B, and S forms of isoenzymes of the j8-o-2-acetamido-2-deoxyhexosidase in human tissues have been accomplished by means of preparative polyacrylamide gel electrophoresis." It was concluded that the A, B, and S forms are composed of a heteropolymer containing a- and j8-chains, a jS-chain homopolymer, and an a-chain homopolymer, respectively. Separation of the isoenzymes of jS-D-2-acetamido-2-deoxyhexosidase in human tissues by electrophoresis on cellulose acetate membranes has been used in the diagnosis of GM -gangliosidosis." The relative abundances and properties of the three isoenzymes were determined, and the relevance of the results to the disease was discussed. 

A rapid method for determining the amylase isoenzymes in human sera and urine is based on separation of the isoenzymes by electrophoresis on a cellulose acetate membrane and visualization using a Blue Starch-agar plate. " The two isoenzymes which are separated possess the same characteristics as salivary and pancreatic amylases. The relative proportions of the isoenzymes are useful in distinguishing hyperamylasemias arising from disorders of the pancreas and parotid gland, etc. 

W5. Windisch, R. M., and Bracken, M. M., Cerebrospinal fluid proteins Concentration by membrane ultrafiltration and fractionation by electrophoresis on cellulose acetate. Clin. Cherru 16,416-419 (1970). 

The support medium provides the matrix in which protein separation takes place. Various types of support media are used in electrophoresis and range from pure buffer solutions in a capfilary to insoluble gels (e.g., sheets, slabs, or columns of starch, agarose, or polyacrylamide), or membranes of cellulose acetate. Gels are cast in a solution of the same buffer to be used in the procedure and may be used in a horizontal or vertical direction. In either case, maximum resolution is achieved if the sample is applied in a very fine starting zone. Separation is based on differences in charge-to-mass ratio of the proteins and, depending on the pore size of the medium, possibly molecular size. 

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