膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | FRET |
| Analyte | RNase A |
| Format | 96T/480T |
| Regulatory Status | RUO |
| Sensitivity | 0.03125 pg |
| Standard Curve Range | 0.03125 pg-2 pg |
| Assay Time | 30 min |
| Suitable Sample Type | For the quantitative determination of RNase A in the environment, some biological materials, common molecular biological reagents such as reaction buffers, enzymes such as reverse transcriptase and RNA polymerase, and buffers for RNA purification and storage. |
| Sample volume | 10 μL & 80 μL |
组分（Materials Provided）
| Items | Size （96tests） | Size（480tests） |
| RNase Substrate | 2 nmol | 10 nmol |
| 10X Reaction Buffer for RNase | 10 mL | 10 mL |
| RNase A (10μg/mL) | 100 μL | 500 μL |
| TE Buffer (pH 7.0) | 1.5 mL | 6 mL |
| Nuclease-free Water | 10 mL | 50 mL |
背景（Background）
RNases are a class of hydrolytic enzymes that catalyzes both the in vivo and in vitro degradation of ribonucleic acid (RNA) molecules into smaller components. RNase enzymes are categorized into two groups: exoribonucleases and endoribonucleases. RNases are ubiquitous in the environment, and in some biological materials, they are present in relatively high concentrations. RNases also frequently contaminate common molecular biological reagents such as reaction buffers, enzymes such as reverse transcriptase and RNA polymerase, and buffers for RNA purification and storage. It is often used DEPC and/or heaten to remove RNases from containers in experiments. Since even only minute amounts of RNase contamination would ruin the experiment, it is necessary to evaluate the presence of RNase with the RNase Activity Assay Kit (Fluorescence).
应用说明（Application）
RNase Activity Assay Kit (Fluorescence) is a convenient and sensitive assay tool to test the presence of RNase in buffers, reagents, and other components.
It is for research use only.
重构方法（Reconstitution）
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储（Storage）
The unopened kit should be stored at -20°C upon receiving.
Find the expiration date on the outside packaging and do not use reagents past their expiration date.
The opened kit should be stored per components table. The shelf life is 3 months from the date of opening.
原理（Assay Principles）
The RNase Activity Assay Kit (Fluorescence) is based on a fluorophore-labeled RNA substrate. When the sample does not contain RNase activity, the substrate is stable and does not produce a fluorescent signal; when the sample contains RNase activity, the substrate is degraded, resulting in a gradual enhanced fluorescence signal, the rate of increase in fluorescence signal is positively correlated with the number and activity of enzymes. Use a fluorescence microplate reader to measure at the wavelength of ex/em= 490/520 nm to determine whether the sample is contaminated by RNase.
活性（Bioactivity）-Fluorescence Please refer to Ds document for the assay protocol.
Add 90 μL RNase Substrate Working Solution (mix RNase Substrate, 10×Reaction Buffer and Nuclease-free Water by 1:1:7 volume) to each 96-well plate, and add 10 μL RNase A standards (0-200 pg/mL×10 μL / well = 0-2 pg/well), incubate the plate in the fluorometer (BMG CLARIOstar) collecting real-time data at one minute intervals for 30 minutes at 37°C using the settings described in this section. The RNase Activity Assay can be evaluated in rigorous kinetic terms using real-time data.
This assay kit employs a standard detection of RNase A. Add 90 μL RNase Substrate Working Solution (mix RNase Substrate, 10×Reaction Buffer and Nuclease-free Water by 1:1:7 volume) to each 96-well plate, and add 10 μL of RNase A standards (0-200 pg/mL×10 μL/well = 0-2 pg/well), incubate for 30 minutes at 37°C. Then measure the fluorescence using the settings described in this section in a fluorometer (BMG CLARIOstar). Take RFU of standards as the ordinate and RNase concentration as the abscissa. Four parameters logistic are used to draw the standard curve. This following data is for reference only.(QC tested)
验证（Validation）
批内差异（Intra-Assay Statistics）
Eight replicates of each of seven samples containing different concentrations of RNase were tested in one assay, Intra-Assay Precision CV＜10%.
批间差异（Inter-Assay Statistics）
Eight replicates of each of seven samples containing different concentrations of RNase were tested in eight independent assays, Inter-Assay Precision CV<15%.
稳定性 (Stability)
The probe is freeze-thawed 3 times, the kit performance meets the requirements, the sensitivity is not reduced, and the CV< 10%.
回收率（Recovery）
Three different concentrations of RNase was spiked into four different kinds of matrixes. The range of the recovery rate is 80%~120%.
干扰效应(Interference effect)
DNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A002) and RNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A001) were used to detect nuclease residues in the same sample. No significant cross-reactivity or interference was observed.
