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The FRET-based assay was adapted to a microarray format and the fluorescence intensity was recorded with a digital camera.
In a first experiment, we were interested to learn whether using the microarray format and calcein-based detection of membrane damage, CTL activity could be specifically detected.
Lastly, we examined expression of the DIAPH3 protein with PCa progression using a cohort of 90 human prostate specimens in a tissue microarray format and a validated anti-DIAPH3 antibody (Supporting Information Fig S2B).
As well as creating assays in the LFA format, it is also possible to print capture antibodies as very small spots in a microarray format and use a comparable assay format to that outlined for NALFIA [ 46].
These results show that P-EIM is sensitive to small molecule-protein interactions and can be used for label-free, high-throughput measurement of proteins in a microarray format and can obtain quantitative molecular interaction kinetic information.
Formalin-fixed paraffin-embedded primary tumor specimens from 289 patients were prepared in a tissue microarray format and used for CHD5 protein expression analysis by immunohistochemical (IHC) staining, which included 19 tumor specimens that were also used for mRNA expression and promoter methylation analyses.
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There have been several protein microarray formats, and each has an advantage depending on what is being detected.
Utilizing RNA data from two independent microarray formats, and immunohistochemical data obtained using antibodies applied in the Human Protein Atlas initiative, we found significant correlations between the RNA and protein profiles of 33% of the gene products.
This technique has been used for detecting prostate-specific antigen [29], as well as simultaneous detection of cytokines' and allergens' specific antibodies in a microarray format [30 32], and introduced commercially for chip-based amplification [20].
Different genome-wide strategies, including different DNA microarrays formats and more recently bisulfite based massive parallel sequencing [8], [9], have been developed to identify genes silenced by CpG hypermethylation in human neoplasias.
Multiple, rapidly evolving fractionation, chemical tagging, mass spectrometry, microarray format affinity methods, and database search algorithms are highlighted; key challenges are much higher throughput, validation of protein identifications, and quantitation.
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