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After rapid drying and cross-linking procedures, the preparation of the membrane array was accomplished.
The membrane array was used to analyze the gene expression of tumor and normal counterpart tissue in 10 CRC patients and 100 peripheral blood specimens.
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A representative picture of the membrane array is shown in Figure 2.
Comparing with glass chip, the current membrane array is relatively simple, time-saving, and the test cost was quite low.
However, the sensitivity of the protein membrane array is very high for CXCL8, ranging from 1 to 25 pg/ml and it thus detects even very small amounts of protein.
Membrane arrays were analysed using ImageJ.
Nylon membrane arrays are typically hybridized with P-dNTP labeled probes and analyzed by a phosphorimager or autoradiography using appropriate densitometric scanning software.
For this analysis, membrane arrays were custom made for by the manufacturer for detection of PDGF-AA, PDGF-BB, IL-13, TNF-α, EGF, MCSCR, insulin-like binding protein 2 (IGFBP-2), IL-1β, HGF, and IL-17.
The membrane bound peptide array was probed with serum of A. fumigatus infected mice for binding according to established procedures [58].
The membrane bound peptide array was probed with the α-85 B antibodies for binding according to established procedures [ 30, 72].
This membrane-bound peptide array was probed with scFv antibody fragments as described above for the immunoblot except that MTT/BCIP [100 µL 1 M MgCl2; 80 µL BCIP (15 g/L in DMF); 120 µL MTT [3- 4,5-dimethylthiazolyl-2 -2,5-diphenyltetrazoliumbromide] (50g/L in 70% DMF +30% H2O2) in 20 mL CBS (8 g/L NaCl, 0.2 g/L KCl, 2.08 g/L citrate, pH 7.0)] was used for staining.
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