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BMC Biotechnol ; 14: 63, 2014 Jul 15.
Article in English | MEDLINE | ID: mdl-25022797

ABSTRACT

BACKGROUND: Inflammatory mediators can serve as biomarkers for the monitoring of the disease progression or prognosis in many conditions. In the present study we introduce an adaptation of a membrane-based technique in which the level of up to 40 cytokines and chemokines can be determined in both human and rodent blood in a semi-quantitative way. The planar assay was modified using the LI-COR (R) detection system (fluorescence based) rather than chemiluminescence and semi-quantitative outcomes were achieved by normalizing the outcomes using the automated exposure settings of the Odyssey readout device. The results were compared to the gold standard assay, namely ELISA. RESULTS: The improved planar assay allowed the detection of a considerably higher number of analytes (n = 30 and n = 5 for fluorescent and chemiluminescent detection, respectively). The improved planar method showed high sensitivity up to 17 pg/ml and a linear correlation of the normalized fluorescence intensity with the results from the ELISA (r = 0.91). CONCLUSIONS: The results show that the membrane-based technique is a semi-quantitative assay that correlates satisfactorily to the gold standard when enhanced by the use of fluorescence and subsequent semi-quantitative analysis. This promising technique can be used to investigate inflammatory profiles in multiple conditions, particularly in studies with constraints in sample sizes and/or budget.


Subject(s)
Cytokines/blood , Enzyme-Linked Immunosorbent Assay/methods , Membranes, Artificial , Animals , Enzyme-Linked Immunosorbent Assay/instrumentation , Humans , Interleukin-1beta/analysis , Luminescent Measurements , Male , Mice , Proteome/analysis , Rats , Rats, Wistar , Tumor Necrosis Factor-alpha/blood
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