We separated proteins of dorsal roots of Macaca Macacus on the basis of differences in their isoelectric points and molecular weights by using a high resolution two-dimensional electrophoresis (2-DE).
The optimal conditions for 2-DE were as follows: The pH gradient for isoelectric focusing (IEF) was prepared by mixing 1 volume of wide range Ampholine, pH 3.5-10, with 4 volumes of narrow range Ampholine, pH 5-7. The composition of gel for IEF of membrane bound proteins was 4%T, 5.4%C, 9M urea, 2% (v/v) Nonidet P-40, and 2% (w/v) Ampholine. The gradient of polyacrylamide for sodium dodecyl sulfate-PAGE was 7-20% T.

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Proteins of nerve roots of Macaca Macacus were separated by using high resolution two-dimensional electrophoresis (2-DE) to identify protein spots specific for either sensory or motor nerve.
The proteins could be separated into 350 spots in soluble fractions and into 520 spots in membrane-bound fraction from both dorsal and ventral roots. Specific protein spots for either ventral or dorsal root could not be detected, however, quantitative differences were observed in some spots (13-16 kDa, pI 6.3-6.6).
Carbonic anhydrase in soluble fraction from dorsal root was identified by immunostaining as heterogeneous spots having different isoelectric points but same molecular weight. On the contrary, the protein spots could not be detected in soluble fractions of ventral root. Carbonic anhydrase should be a marker protein to distinguish the sensory nerve from the motor nerve.

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Serum amyloid A (SAA) protein is a sensitive and rapid acute phase reactant and its meaurement is gaining wide spread acceptance for the monitoring of inflammatry diseases and assessing the prognosis in secondary amyloidosis.
We set up a latex agglutination nephelometric immunoassay system for the quantitation of SAA in human serum. SAA-enriched HDL (high density lipoprotein) which SAA content was determined electrophoretically, was used as assay standard. The values determined by this method related with enzyme immunoassay using the same antibody and standard. The mean value of SAA in normal serum (n=100) was 1.1μg/ml and ranging 0.48-4.8μg/ml (mean±2 SD).
The log SAA vs log CRP ratio showed essentialy the same values in patients with rheumatoid arthritis, acute myocardial infarction, bacterial pneumonia, lung cancer, gastrointestinal cancer, surgical operation, while significantly higher values were obtained in patients with rubellosis, cytomegalo virus infection and kidney allograft rejection. This rapid measurement method of SAA may fit application for the routine use.

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Mouse liver ribosomes were analyzed by 1.8% polyacrylamide 0.5% agarose composite gel electrophoresis (7.5% cross linkage gel). When the ribosomal suspension mixed with agarose was applied to the gel slot, ribosomes were resolved in the composite gel. When turned gel electrophoresis in which gel concentration was the same for the second run, was carride out, ribosome bands from 40S subunits to the maximum hexamers were observed. This high resolution electrophoresis may be one of the most powerful tools available to analyze the non-denatured ribonucleoprotein particles and membrane components and to study the conformational change of ribosomes.

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A novel 36kDa rhamnose-binding lectin isolated from Plecoglossus altivelis roe (PA 36), a probe for L-rhamnose and α-D-galactosyl nonreducing termini, bound to and agglutinated murine tumor cells rather than human type B erythrocytes. PA 36 was purified by DEAE-cellulose ion exchange and rhamnose-Sepharose affinity chromatography. The most effective saccharide in the agglutination inhibition assay was L-rhamnose. The α-D-galactosyl containing saccharides, such as melibiose, raffinose and stachyose, were more effective than the β-D-galactosyl containing saccharide, lactose. Lectin binding glycoproteins were determined by SDS-PAGE and blotting, followed by reaction with horseradish peroxidase-labeled PA 36. The glycoproteins ranging in molecular weight from 66, 000 to 72, 000 and deriving from murine tumor cells and bands 4.1 and 4.2 deriving from human type B erythrocytes were PA 36-binding glycoproteins. These results raise the possibility that nonreducing terminal α-D-galactosyl-containing glycoproteins composed of heterogeneous carbohydrate chains may be present on the surface of a number of different murine tumor cells as well as on human type B erythrocytes.

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An investigation was conducted to determine the most appropriate conditions for analysis of single-strand conformation polymorphism (SSCP) in the typing of HLA-DQA gene segments after PCR amplification. The original SSCP conditions (conformation of DNA fragments in 95% formamide and separation by electrophoresis on a gel containing 10% glycerol) were modified so that the same reagents as those used at the conformation and electrophoresis stages. could be applied for the SSCP analysis. Three reagents (10% glycerol, 10% formamide and 5% saccharose) were tested. It was found that all modifications gave more distinct band images and separation than the original technique. The clearest images were obtained when a 5% saccharose aliquot was used, whereas the most sensitive discrimination of SSCP bands in terms of Rf was achieved using glycerol.

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