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제품 설명
Salt Active UltraNuclease ELISA Kit
제품 번호
36719ES48 / size: 48T-EN
36719ES96 / size: 96T-EN
제품 설명
Salt Active UltraNuclease is a non-specific endonuclease which derived from Marine microorganisms and is expressed and purified in Escherichia coli (E. coli) by genetically engineered. This nonspecific, recombinant endonuclease has optimum activity at high salt concentration (500mM NaCl), which can improve efficiency and yield in various workflows.
This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residual Salt Active UltraNuclease in samples. First, add the Salt Active UltraNuclease standard and test sample to the Anti-Salt Active UltraNuclease coated microtiter strips (36719-A), then add the diluted biotin labeled Anti-UltraNuclease antibody (36719-C), and finally add Streptavidin-HRP (36719-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, add TMB substrate (36719-H) into the complex to observe color reaction after washing the complex. TMB is converted into blue under the catalysis of HRP enzyme and finally converted into yellow in the presence of acid, and the shade of color is positively correlated with the amount of Salt Active UltraNuclease in the sample.
The detection range of this kit is 0.047-3 ng/mL; the lower detection limit is 0.024 ng/mL.
Sensitivity | 0.024 ng/mL (range 0.047~3 ng/mL) |
Assay Time | <4 hours |
Assay Principle | Two-site immunoenzymetric assay |
Signal Amplification | Biotin-Streptavidin system |
Detection Wavelength | 450nm-630nm |
Components No. | Name | 36719ES48 | 36719ES96 |
36719-A | Anti-Salt Active UltraNuclease coated microtiter strips | 48 T | 96 T |
36719-B* | Standard: Salt Active UltraNuclease | 1 vial | 2 vial |
36719-C | Detection Antibody：Biotin-conjugated Antibodies | 60 μL | 120 μL |
36719-D | Streptavidin-HRP | 30 μL | 60 μL |
36719-E | Dilution Buffer 1 | 25 mL | 50 mL |
36719-F | Wash Buffer Concentrate (20×) | 25 mL | 50 mL |
36719-G | Dilution Buffer 2 | 15 mL | 30 mL |
36719-H | TMB Substrate | 8 mL | 15 mL |
36719-I | Plate Sealer | 3 each | 5 each |
* 1. The stop solution is not provided in this kit, it need to be prepared by user. 1M HCl can be used as the stop solution;
* 2. The Standard (36719-B) is Lyophilized powder.
This product should be stored at 2°C ~8°C. Unopened product is valid for one year. Once the reagent is opened, it is valid for half a year.
*Upon receipt of the kit, please check whether all components are complete and immediately store them in corresponding condition.
Figures 1. The specificity of Salt Active UltraNuclease ELISA kit was shown in the figure above
7 samples were estimated for the non-specific reaction with the antibodies in the kit. The concentration of the 7 samples were all below the QL (0.047ng/mL).
A）Repeatability
Spiked sample | Intra-Assay | ||
Duplicates | Average Conc. (ng/mL) | CV (%) | |
S-1.5 ng/mL | 10 | 1.344 | 2.97% |
S-0.375 ng/mL | 10 | 0.260 | 2.27% |
S-0.094 ng/mL | 10 | 0.061 | 2.56% |
B）Intermediate precision
Spiked sample | Inter-Assay | ||
Duplicates | Average Conc. (ng/mL) | CV (%) | |
S-1.5 ng/mL | 30 | 1.335 | 5.70% |
S-0.375 ng/mL | 30 | 0.258 | 5.59% |
S-0.094 ng/mL | 30 | 0.064 | 6.82% |
Figures 2. The precision of Salt Active UltraNuclease ELISA was assayed as the above scheme
10 duplicated were tested at 3 concentrations on the standard curve, and all the CV% were lower than 5% (Figure 2A). 3 lot of Salt Active UltraNuclease ELISA kits were tested and the CV% were lower than 10% (Figure 2B).
Dilution factor | Dilution recovery (%) | Range of Dilution recovery（%） |
1:2 | 92.5 | 87.5-98.9 |
1:4 | 87.0 | 79.0-95.2 |
1:8 | 92.6 | 78.9-106.2 |
1:16 | 96.9 | 79.1-118.1 |
Spiked samples | Spike recovery (%) | Range of Spike recovery（%） |
S-0.75ng/mL | 105.3 | 91.0-122.1 |
S-0.18ng/mL | 96.6 | 86.5-105.5 |
Figures 3. The accuracy of UCF.ME™ UltraNuclease ELISA was assayed as the following scheme
The protein sample with high Salt Active UltraNuclease concentration was serial 2 fold diluted and also spiked with 0.75ng/mL and 0.18ng/mL Salt Active UltraNuclease. The dilution recovery and the spike recovery were all within 75%-125% (Figures 3).
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