Electrophoresis alkaline

The enzymatic radioassay method for the analysis of acetylcholine and choline in brain tissue has been reported by Reid et al. [210]. The method describes the determination of nanogram amounts of acetylcholine and choline in as little as 10 mg of brain tissue, involves isolation of acetylcholine by high-voltage paper electrophoresis, alkaline hydrolysis of acetylcholine to choline, and conversion of this into [32P]-phosphoryl choline in the presence of choline kinase and [y32P] ATP. The labeled derivative is isolated by column chromatography on Bio-Rad AG1-X8 resin, using Tris buffer solution as the eluent. Cerenkov radiation from 32P is counted (at 33% efficiency) in a liquid scintillation spectrometer. The amount of phosphorylcholine is proportional to the amount of choline over the range of 0.08-8.25 nmol. 

Biomedical Applications. TRIS AMINO is used for a number of purposes in its pure form, it is an acidimetric standard the USP grade can be utilized intraveneously for therapeutic control of blood acidosis TRIS AMINO also is useful in genetic engineering as a buffering agent for enzyme systems, industrial protein purification, and electrophoresis. AMP has found use as a reagent in enzyme-linked immunoassays. The primary appHcation is for alkaline phosphatase assays. 

Gasiorowski, K., Brocos, B. DNA repair of hydrogen peroxide-induced damage in human lymphocytes in the presence of four antimutagens. A study with alkaline single cell gel electrophoresis (comet assay). Cellular Molecular Biology Letters, Vol.6, (2001), pp. 897-911, ISSN 1425-8153... 

With the development of HPLC, a new dimension was added to the tools available for the study of natural products. HPLC is ideally suited to the analysis of non-volatile, sensitive compounds frequently found in biological systems. Unlike other available separation techniques such as TLC and electrophoresis, HPLC methods provide both qualitative and quantitative data and can be easily automated. The basis for the HPLC method for the PSP toxins was established in the late 1970 s when Buckley et al. (2) reported the post-column derivatization of the PSP toxins based on an alkaline oxidation reaction described by Bates and Rapoport (3). Based on this foundation, a series of investigations were conducted to develop a rapid, efficient HPLC method to detect the multiple toxins involved in PSP. Originally, a variety of silica-based, bonded stationary phases were utilized with a low-pressure post-column reaction system (PCRS) (4,5), Later, with improvements in toxin separation mechanisms and the utilization of a high efficiency PCRS, a... 

Free a-and 3-chains can be demonstrated In a freshly prepared hemolysate using starch gel electrophoresis at alkaline pH. 

The presence of Individual chains In a hemoglobin variant can also be demonstrated by electrophoresis at alkaline pH after the protein has been dissociated Into Its subunits through exposure to 6 M urea In the presence of 3-mercaptoethanol. The buffer is either a barbital buffer or a tris-EDTA-boric acid buffer, pH 8.0 - 8.6, and contains 6 M urea and 3-niercapto-ethanol. Dissociation of the hemoglobin Into subunits Is best accomplished In a mixture of 1 ml 10 g% Hb (or whole hemolysate), 4 ml 6 M urea barbital or tris-EDTA-boric acid buffer, and 1 to 1.5 ml 3-mercaptoethanol. After 30 minutes to 1 hour the sample Is subjected to cellulose acetate or starch gel electrophoresis. Each chain has a specific mobility and an alteration In electrophoretic mobility easily Identifies the abnormal chain. 

Alkaline phosphatase assays based on 3-glycerophosphate now appears to be obsolete, and methods buffered by either glycine or barbital are also obsolete as these buffers inhibit ALP or are poor buffers. Serum alkaline phosphatase is known to be composed of several isoenzymes which presumably arise from bone, liver, intestine, and placenta. The placental alkaline phosphatase is known to be much more resistant to heat denaturation than the other isoenzymes, and this resistance provides a simple test for it (5). The other enzymes can be separated through the differential inhibition by phenylalanine, by electrophoresis and by specific antibodies. However, the clinical usefulness of the results obtained is in doubt (23). 

Bachmartn, K., Boden, J., and Haumann, I., Indirect fluorimetric detection of alkali and alkaline earth metal ions in capillary zone electrophoresis with cerium (III) as carrier electrolyte, /. Chromatogr., 626, 259, 1992. 

A variety of buffers is used in electrophoresis. The selected buffer must contain ions to carry the current. Other than current-carrying capacity, the most critical criterion for buffer selection is the stability of the sample to be analyzed. Many proteins are unstable in acidic pHs, so alkaline buffers are frequently employed. Tris-(hydroxymethyl)amino methane (TRIS or THAM), sodium acetate, and ethylenedi-aminetetraacetate (EDTA) are common solutes in buffers, with pHs between 7.9 and 8.9 typical. (Refer to Chapter 5 for a discussion of buffers.) These buffers also work well with nucleic acid fragments. In addition, phosphate buffers, e.g., 10 mMK3P04, are often used with nucleic acid fragments (1.0 mM = 0.0010 M). 

Sengelpv, H., Nielson, M. H., Borregaard, N. (1992). Separation of human neutrophil plasma membrane from vesicles containing alkaline phosphatase and NADPH oxidase activity by free flow electrophoresis. J. Biol. Chem. 267,14912-17. 

Sample preparation. All allantoic fluid of chicken embryos or calf serum used in experiments contained influenza virus (104—106 EID50/ml). The samples of biological fluids underwent photodynamic treatment as described above. One milliliter aliquots were taken before treatment and at 3 and 6 h after the start of experiment. To analyze the effect of photodynamic treatment on proteins we used alkaline denaturing electrophoresis in the presence of sodium dodecylsulfate (SDS) and P-mercaptoethanol (P-ME). 

Carvalho has studied the separation of technetium from rhenium by electrophoresis and paper chromatography. Good separations can be obtained by electrophoresis using either alkaline hydra2dne sulfate or a solution of SnClj in hydrochloric acid as electrolytes. Both elements can sufficiently be separated by paper... 

Alkaliphilic Bacillus sp. AM-001 was aerobically grown to the early stationary phase at 37°C in alkaline medium (pH 9.0) containing 1% konjak mannan. Total chromosomal DNA obtained by the method of Saito and Miura( 14) was digested with Hindlll restriction enzyme. And 2 to 4 kbp DNA fraction of chromosomal DNA was collected by 1% agarose gel electrophoresis. The plasmid pUC19 was digested with Hindlll and then dephosphorylated with bacterial alkaline phosphatase. After the... 

Sardas S, Aygun N, Gamli M, et al Use of alkaline comet assay (single cell gel electrophoresis technique) to detect DNA damage in lymphocytes of operating room personnel occupationally exposed to anaesthetic gases. Mutat Res 418(2-3) 93-100, 1998... 

NT044 Tsuda, S., N. Matsusaka, S. Ueno, N. Susa, and Y. F. Sasaki. The influence of antioxidants on cigarette smoke-induced DNA single-strand breaks in mouse organs a preliminary study with the alkaline single cell gel electrophoresis assay. Toxicol Sci 2000 54(1) 104-109. 

For the first two entries, alkaline sucrose gradients were used all others employed gel electrophoresis. Degree of hydration, deduced from known relative humidity, is given as r, the molar ratio of water to mononucleotide. Sample temperature during radiation. 

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