Electrophoresis documentation

Reference data on common chitosan salts are useful to assess the characteristics of complexes made by modified chitosans and DNA. Complexes formulated with several chitosan salts were assessed for complexing ability toward DNA, transfection efficiency in Chinese hamster ovary cells, and their effect on cell viability. Chitosan hydrochloride, lactate, acetate, aspartate and glutamate (chitosan MW 45 kDa) formed complexes with pcDNA3-CMV-Luc. Gel electrophoresis documented that complexes formed at N/P ratios above 2 with all chitosan salts. The transfection efficiency depended on the anion, on the chitosan MW, and on the N/P ratio maximum transfection efficiencies were at N/P ratio of 12, 12, 8, 6 and 6 for said salts, respectively. The MTT assay indicated that all chitosan/DNA complexes had low cytotoxicity (Weecharangsan et al., 2008). 

The entire chapter <(1047) is harmonized with the Japanese and the European Pharmaceopoeias. In order to have a parallel organization of the harmonized documents, the current chapter <( 1047) will be divided and replaced by six general information chapters, including <(1053) Biotechnology Derived Articles — Capillary Electrophoresis. 

After electrophoresis, transfer the gel into ethidium bromide solution (50 pi stock solution in 1 1 of tap water) and stain for 5 -10 min. Transfer the gel onto a transilluminator and visualise the DNA with UV light, taking care to protect the skin and eyes against UV radiation. Document the result with a standard photograph or digital image analyser. 

As a rule, chemical methods used in the examination of writing materials require initial preparation of a sample for study. Paper chromatography, thin-layer chromatography and capillary electrophoresis are experimental techniques often applied. These methods lead primarily to separation of the dyes contained in the ink under examination and to the discrimination of ink samples. The techniques are simple to use, require a small amount of sample for examination, are selective and give reproducible results. Their basic disadvantage, however, is the necessity to isolate the ink from the substrate (e.g. paper) on which the examined document has been prepared. Solvent extraction of the ink often leads to partial damage of the document. 

The presence of peptides and amino acids in gastric juice was demonstrated many years ago by Nencki and Sieber (N3), Babkin (Bl), and Komarov (K22). This became better documented only more recently with the use of modem physicochemical tools, such as paper partition chromatography, electrophoresis, and polarography. Demonstration of the products of proteolytic degradation of serum proteins and muco-substances has also been made possible only by the use of these methods. The reader is referred to the second article by the author (G21) for pertinent information and to the article by Bouda (B29). 

The usual technique for identification of AChE is fractionation by polyacrylamide gel electrophoresis followed by incubation of the gel with the substrate acetylcholine and copper ions. AChE migrates more rapidly than PChE and appears as a distinct white precipitate band below the PChE band. That the second band is AChE Is documented by inhibition of the enzyme activity in the band by the specific AChE inhibitor l,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide—BW284C51. No method is avail-able commercially laboratories develop their own assay systems using published methodology. Enzyme immunoassay using a monoclonal antibody specific for neurally derived AChE has also been used. ... 

The results obtained from urea gradient gel electrophoresis experiments compare favorably with what has been reported in the literature for the unfolding/folding of carbonic anhydrase. The existence of an intermediate species at 1.2-2M GuHCl or 5M urea has been documented (11,14,16,26,36,37),... 

Documentations obtained when evaluation is performed with a flat-bed scanner are always dependent on the software used and also, of course, on the scanned object. As mentioned in Section 7.4, a flat-bed scanner can be directly used for processing colored chromatograms only. The first investigations were done with the Pharmacia system, which was also used for the evaluation of DNA electrophoresis gels. 

Use of Complex Standards for Cell Proteins and Two-Dimensional Electrophoresis. One of the more widely-used cell lines chosen as a reference standard is the lymphoblastoid cell line GM607, derived from a normal individual and available from the Human Genetic Mutant Cell Depository, Camden, NJ 08103. This cell line may be grown in defined media, labeled with a radioactive tracer, and reproducibly separated in a 2-DE system. Heat shock proteins may readily be isolated and visualized from this cell line, as shown by Anderson et al. (42). For serum, a reference preparation for serum proteins is available as a certified reference material prepared and assayed by the College of American Pathologists (CAP) and by the U. S. Centers for Disease Control. A widely available human serum standard is that provided by the National Bureau of Standards as SRM 909. If sufficient interest from the user community is evident, a full electrophoretic characterization of this material can be included in the documentation. If the amount of selected standard proteins loaded on a gel is known, "relative" quantification of similar proteins could be obtained. In addition, the National Bureau of Standards could serve as an impartial evaluator of potential national standards (e.g. molecular weight standards, "tie-point" proteins, and Isoelectric point standards) to assess suitability and stability. 

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