- Sensitive - detect as little as 3.5 x 10-7 units (~0.5 pg) of RNase A
- Simple, straightforward 30 min assay
- Detect with a fluorometer or by eye
- Monitor RNase activity in column fractions during protein purification
- Use with DNaseAlert™ to quantitatively detect both RNases and DNases in the same sample
If you need to detect RNases, then RNaseAlert is the kit of choice. This patent-pending technology reports RNase activity in a convenient and sensitive assay that delivers fast, accurate results in real-time. The RNaseAlert Kit uses a novel RNA substrate tagged with a fluorescent reporter molecule (fluor) on one end and a quencher on the other. In the absence of RNases, the physical proximity of the quencher dampens fluorescence from the fluor to extremely low levels. When RNases are present, however, the RNA substrate is cleaved and the fluor and quencher are spatially separated in solution. This causes the fluor to emit a bright green signal when excited by light of the appropriate wavelength. Fluorescence can be readily detected by eye upon illumination on a UV box, or with a filter-based or monochromator-based fluorometer. Since the fluorescence of the RNaseAlert Substrate increases over time when RNase activity is present, results monitored with a fluorometer can be evaluated kinetically. The sequence of the RNaseAlert Substrate has been carefully optimized to detect several RNases, including RNase A, RNase T1, RNase I, micrococcal nuclease, S1 nuclease, mung bean nuclease, and Benzonase®.
Rapid and Convenient Protocol
The RNaseAlert procedure requires just minutes to setup. The lyophilized, fluorescent substrate is reconstituted in the supplied tube and incubated with the test solution at 37°C for 30-60 min. The results are then read on a transilluminator or fluorometer. Samples that noticeably fluoresce compared to the negative control are contaminated and should not be used with RNA. Nuclease-free water is provided as a minus-RNase control, and RNase A is included as a plus-RNase control. Solutions that fail to fluoresce are safe to use with RNA, although testing should be repeated periodically or whenever RNase contamination is suspected.
High Throughput and Lab Test Kits Available
RNaseAlert is provided in two convenient kit formats. The RNaseAlert Lab Test Kit (see schematic) is suitable for testing small sample numbers and can be used to ensure that solutions, tubes, tips, etc. are RNase-free. The RNaseAlert QC System (see schematic) is designed for high-throughput assays in a 96-well format. This kit contains enough substrate to test 480 samples (5 x 96) using a fluorometer. The superior sensitivity of fluorometer measurements allows multiple contaminated samples to be identified in minutes, rather than the 30-60 minute incubation recommended for readout using a transilluminator.
Quantify RNase & DNase Activities in Real-time Simultaneously
RNaseAlert is also fully compatible with Ambion's DNaseAlert™ QC System. Since the DNaseAlert Substrate contains a fluorescent tag that is spectrally distinct from RNaseAlert, the two kits can be used simultaneously with a fluorometer for quantitative real-time analyses of both RNases and DNases within a single sample.
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