A simple and economical procedure for liquid scintillation counting of tritiated compounds on glass-fibre filter discs

doi:10.1016/0020-708X(80)90001-0

The International Journal of Applied Radiation and Isotopes

Volume 31, Issue 11, November 1980, Pages 665-669

https://doi.org/10.1016/0020-708X(80)90001-0 Get rights and content

Abstract

A simple and economical procedure has been set up for the liquid scintillation counting of both soluble (aqueous solutions of pure tritiated thymidine) and insoluble (tritiated culture pellets of stimulated or unstimulated lymphocytes) compounds on glass-fibre filter discs. It has been found that the radioactivity of both the soluble and insoluble compounds can be detected in only 0.1 ml of scintillation cocktail (aqueous counting scintillant, ACS). The addition of larger amounts of scintillator (0.1–3 ml) to vials containing insoluble compounds in no way increases the counting rate. With soluble compounds (previously dried at 100°C), increased quantities of scintillator (0.1–1 ml) yield slightly higher counts, but with volumes of 2 or 3 ml the counts remain stable.

Résumé

Un procédé simple et économique a été réalisé pour compter dans un très faible volume de liquide scintillant des composés solubles (solution acqueuse de thymidine tritiée) ou insolubles (culot radioactif de culture de lymphocytes stimulés ou non) déposés sur les disques en fibre de verre.

Les résultats montrent que la radioactivité de chacun de ces composés peut être détectée en utilisant seulement 0,1 ml de liquide scintillant (aqueous counting scintillant-ACS). L'addition de quantités croissantes de scintillant (0,1–3 ml) dans des fioles contenant des composés insolubles ne provoque aucune augmentation du taux de comptage, tandis que l'addition du même scintillant (0,1–1 ml) à des composés solubles induit une légère augmentation du taux de comptage. La radioactivité reste pratiquement indentique avec 2 ou 3 ml de scintillant.

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There are more references available in the full text version of this article.

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Rice lectin (Oryza sativa, var. Balilla 28) was purified from defatted embryos by aqueous acid extraction at pH 1.3 followed by ammonium sulfate precipitation between 2 and 4 m, affinity chromatography on agarose-p-aminophenyl-β-d-N-acetylglucosamine,and gel filtration on AcA 54. The homogeneity of the lectin was checked by polyacrylamide gel electrophoresis, gel filtration, and immunodiffusion. The amino acid analysis revealed a high half-cystine content (9%) and a low aromatic and hydrophobic amino acid content. The lectin contained neither neutral carbohydrates nor amino sugars. The isoelectric point was estimated to be 8.1. The molecular weight of rice lectin was estimated to be 38,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions showed two polypeptides with M_r 19,000 and 15,000. The circular dichroism spectrum of rice lectin in far ultraviolet was characterized by a positive maximum at 228 nm and a negative band at 203 nm suggesting the presence of a β-pleated sheet and the absence of α-helix. Rice lectin had no human blood group specificity and agglutinated rabbit erythrocytes more efficiently than erythrocytes from other animal species. Furthermore, agglutination was enhanced by trypsin treatment of erythrocytes. The erythroagglutinating activity was very high since the minimal concentration needed to agglutinate erythrocytes was 0.05 μg/ml. Although [methyl-³H]thymidine incorporation was stimulated in human lymphocytes, rice lectin could not be considered as a mitogenic lectin since it stimulated neither blast transformation nor lymphocyte proliferation. The saccharide specificity of rice lectin was related to N-acetylglucosamine and its oligomers: N,N′,N″-triacetylchitotriose was the most powerful inhibitor. Furthermore, the N-acetylneuraminic acid was not a specific rice lectin determinant. Finally, the double immunodiffusion method revealed a cross-reactivity between rice lectin and wheat germ agglutinin, indicating that these lectins were closely antigenically related. The analogies and differences between biological and immunological properties of rice lectin and wheat germ agglutinin are discussed and the possibility of their evolution from a common ancestor is put forward.
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The International Journal of Applied Radiation and Isotopes

A simple and economical procedure for liquid scintillation counting of tritiated compounds on glass-fibre filter discsUne Méthode Simple et Economique pour Compter en Scintillation Liquide des Composés Tritiés, Fixés sur des Filters en Fibre de Verre

Abstract

Résumé

Biochim. biophys. Acta

Cell. Immun.

Isolation, molecular and biological properties of a lectin from rice embryo: Relationship with wheat germ agglutinin properties

Calmodulin in lymphocyte mitogenic stimulation and in lymphoid cell line growth

Stimulation of 2′, 3′-cyclic nucleotide 3′-phosphodiesterase in human lymphocytes by robinia pseudoacacia lectin

Use of pliable bags in liquid scintillation counting

A microquantitative method for investigating the interaction between labeled lectins and the surface membranes of human lymphocytes using a semi-automatic harvesting machine