Optimized Blocking Of Porous Nitrocellulose Films For Sensitive Protein Microarrays
| Autor: | Michael A. Shultz, Aki Ohdera, Jason MacManiman, Charles M. McGrath |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 54, Iss 4, Pp 223-225 (2013) |
| Druh dokumentu: | article |
| ISSN: | 00011401 1940-9818 0736-6205 |
| DOI: | 10.2144/000114011 |
| Popis: | Abstract Super G Blocking Buffer was developed to produce low backgrounds in assays using nitrocellulose film-slides. Using this blocking reagent in combination with ONCYTE® SuperNOVA Film-Slides, we achieved an order of magnitude greater sensitivity in a cytokine microarray assay than with hydrogel- or silane-based glass substrates. Limit of detection (LOD) ranged from 160 – 690 fM for IL1α, IL1β, IL6, TNFα, TNFβ, and IFNγ without use of enzymatic signal amplification. The dynamic range for these microarrays on Film-Slides was greater than that of hydrogels and functionalized glass substrates and the LOD was 50-fold lower. On Film-Slides, the linear dynamic range spanned 6 orders of magnitude and the LOD was pushed into the zmole range (62.5 zmol). The advantages of Film-Slides are discussed in the larger context of developing the most sensitive protein microarrays for multiplexed clinical assays and quantitative studies of the cellular proteome. |
| Databáze: | Directory of Open Access Journals |
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