Protein Determination by UV Absorption

-An efficient antioxidant system is of particular importance for insects whose high metabolic rates promote increased formation of reactive oxygen species (ROS). The amount of ROS can be additionally increased by environmental factors. This study investigates the ability of red mason bees (Osmia bicornis L.) to inactivate free radicals during insect development. Both male and female bees were studied, from the larval to the active imago stage. The activity of four antioxidant enzymes, superoxide dismutase, catalase, peroxidase and glutathione Stransferase, was measured; and glutathione content and total antioxidant status were determined. The highest values of the examined parameters were found in feeding stages-in larvae and in active imagines of both genders. Significant differences between genders were noted mainly in catalase activity, which was lower in overwintering imagines and active females than in males. Most differences were observed between females and males after emergence.antioxidants / pollinating insects / insect metabolism / development / enzymes

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“…The proteins were assayed spectrophotometrically at a wavelength of 280 nm (Aitken and Learmonth 1996).…”

Section: Methodsmentioning

confidence: 99%

Variations in antioxidant defense during the development of the solitary bee Osmia bicornis

et al. 2014

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“…The bound proteins were eluted using 0.2 M Glycine HCl (pH 2.2) and the pH of the eluted fractions was brought to neutral by adding 2 M Tris. The absorbance of fractions was measured at 260 and 280 nm on a spectrophotometer (Perkin Elmer Lambda 25) and the protein concentration was estimated (Aiken and Learmoth 1996). The column was regenerated using 0.1 M Tris-HCl, 0.5 M NaCl, pH 8.5, 0.1 M Sodium acetate and 0.5 M NaCl, pH 4.5 after each use.…”

Section: Construction Of Immunoaffinity Columnmentioning

confidence: 99%

“…The eluted bound protein was dialysed extensively against 20 mM tris-saline buffer, pH 7.4, concentrated with PEG 20,000 and was designated as affinity purified P. epiclitum somatic antigen (Aff-PSAg). The protein content of the antigen was determined spectrophotometrically (Aiken and Learmoth 1996).…”

Section: Construction Of Immunoaffinity Columnmentioning

confidence: 99%

Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum

et al. 2010

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Polypeptide profile of somatic antigen of Paramphistomum epiclitum (PSAg) and Gastrothylax crumenifer (GSAg) was studied by SDS-PAGE. PSAg and GSAg showed 14 and 19 polypeptides in the range of 14.9-95.5 and 13.7-129.6 kDa with six common polypeptides of mol wt 16.8, 21.8, 23.7, 35.5, 43.4 and 70.8 kDa. P. epiclitum experimentally infected sheep sera were used for identification of specific immuno-dominant peptide in the range of 37-40 kDa against P. epiclitum by western blotting. Hyperimmune sera (HIS) was raised in rabbit against the identified polypeptide, IgG was separated from HIS and an immunoaffinity column was constructed with a binding percentage of 83.74 of IgG with CNBr activated Sepharose 4B. Purification of somatic antigen (PSAg) was done with immunoaffinity chromatography and 37-40 kDa protein antigen was isolated in pure form with recovery percentage of 2.97%. This purified fraction of somatic antigen can be used as a candidate antigen for development of serological assay for early diagnosis of paramphistomosis among livestock.

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“…Protein content was determined by the spectrophotometric method at 280 nm wavelength (Aitken & Learmonth 1996). The result obtained was expressed as mg of protein contained in ml of hemolymph.…”

Section: Protein Contentmentioning

confidence: 99%

Total protein and carbohydrate content and protease and disaccharidase activities in the hemolymph of Lymnaea stagnalis naturally infected with digenean larvae

et al. 2013

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Lymnaea stagnalis is an intermediate host of many Digenea. The infestation affects host metabolism. The aim of the work was to investigate hemolymph biochemical indicators of L. stagnalis infected with four species of trematodes: Diplostomum pseudospathaceum, Paryphostomum radiatum, Plagiorchis elegans or Opisthioglyphe ranae. The protein profiles and proteinase activity in the hemolymph of sexually mature individuals of Lymnaea stagnalis maintained at 19 • C were tested. As the carbohydrates are main substrates for energetic metabolism of the great pond snail their content and disaccharidase activity were also studied. Hemolymph samples were collected during weeks 3 and 4 of rearing. No significant differences in the total protein content between uninfected individuals and snails infected with the first three trematode species were detected. In the snails infected with O. ranae the quantity of total proteins was near twice higher than in those uninfected. A higher share of 70 kDa proteins in infected than in uninfected snails as well as reduction of the low molecular weight fractions of proteins for snails infected with D. pseudospathaceum and P. radiatum were detected. During week 3, carbohydrate content in the infected snails did not differ from that in the controls while during week 4 it was significantly lower in snails infected with P. elegans or O. ranae. The content of the major soluble carbohydrate in the hemolymphsaccharose -changed in a similar way. No activity of trypsin or pepsin in the hemolymph sample was detected while the activity of chymotrypsin was lower in infected snails vs. controls. On the other hand, saccharase and maltase activities were higher in infected than in uninfected snails. The biochemical hemolymph indicators in naturally infected host-snails show some differences depending on the parasite species but they are not sufficiently species-specific to offer the basis for establishing the model unique for a particular parasitosis. Our results from the field did not always coincide with those from the laboratory.

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Protein Determination by UV Absorption

Cited by 121 publications

References 7 publications

Variations in antioxidant defense during the development of the solitary bee Osmia bicornis

Variations in antioxidant defense during the development of the solitary bee Osmia bicornis

Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum

Total protein and carbohydrate content and protease and disaccharidase activities in the hemolymph of Lymnaea stagnalis naturally infected with digenean larvae

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