Dynamic range of protein binding for ONCYTE microporous film slides, PATH thin film nitrocellulose slides, and competing nitrocellulose and hydrogel slides. Data are normalized, background-subtracted relative fluorescence intensities of spotted TRITC-labeled IgG collected at 532 nm. All slides were blocked (with an appropriate blocking buffer for each slide type) and washed to remove unbound protein. Dynamic range of protein binding spans over 7 orders of magnitude with a linear range of 6 orders of magnitude (r2 = 0.999) for ONCYTE and PATH films. Note the short dynamic range of the hydrogel slides due to poor protein binding at lower protein concentrations as well as limited protein binding capacity. Note also the truncated dynamic range for the competing porous nitrocellulose film slide due to high intrinsic autofluorescence from that manufacturer. Data presented are the mean ± standard deviation for N = 4 slides per slide type.
ONCYTE® Brand Nitrocellulose Thin Film Slides
|Dynamic Range (log scale fluorescence||5-6||5-6||7+||4-5|
|Low lot-to-lot Variation
|Hydrphobicity1 (may affect spot size)||+||+||++||++|
|Best for any application requiring high binding capacity and colorimetric detection.||Reduced fluorescence background with lower binding capacity than AVID. Good signal-to-noise ratio for fluorescence detection.||Second generation NOVA, lowest fluorescence background, high binding capacity. Best for fluorescence detection and large dynamic range.||Lowest fluorescence background, lower binding capacity, reduced dynamic range. Best signal-to-noise ratio for fluorescence detection.|
1. More hydrophobic surfaces may result in reduced spot diameter, depending on the spotting buffer composition.Results may vary based on buffers, sample preparation, spotting and scanning instruments. See our Oncyte® user guide for full discussion