This kit is suitable for the quantitation of Nonspecific Endonuclease (SMNE) in purification process of bioproducts. It can detect genetically engineered endonucleases from Serratia marcescens, such as Benzonase® endonuclease.
This SHENTEK® Nonspecific Endonuclease (SMNE) ELISA Kit is based on the solid-phase enzyme-linked immunosorbent assay (ELISA) with a double-antibody sandwich technique to detect residual nonspecific endonuclease (SMNE). A sheep polyclonal antibody specific to SMNE was employed in the assay to capture any remaining SMNE impurities in the sample. Both the Calibration standards and test samples were simultaneously added to the microtiter plate coated with affinity purified capture antibody and followed by incubation and washing. The biotinylated antibody was added to the microtiter plate to bind the SMNE and then reacted with streptavidin labeled with HRP (Horseradish Peroxidase). TMB (3,3',5,5' -tetramethylbenzidine) substrate was added into reaction, HRP catalyzed the oxidation of TMB by H2O2 to produce a blue product (maximum absorption peak at 655 nm). Then the stop solution was added to terminate the enzymatic reaction, resulting in a yellow colored product (maximum absorption peak at 450 nm). The absorbance values at 450 nm wavelength were positively correlated with the SMNE concentration in the Calibration standard and the sample. The concentration of SMNE in the sample can be calculated using a dose-response curve.
| Components | Shipping Condition | Unit Size | Detection Method |
| SMNE Calibration Standard | 2-8℃ | 96 Tests | Sandwich ELISA |
| Anti-SMNE Microtiter Strips | |||
| SMNE Diluent | |||
| Wash Buffer Concentrate (10×) | |||
| Anti-SMNE:Biotinylated Conjugate (100×) | |||
| Streptavidin-HRP (100×) | |||
| TMB Substrate | |||
| Stop Solution | |||
| Sealing Film |
| Linearity & Range | 0.25-25 ng/mL, R2>0.990 |
| Accuracy | Recovery Rate= 96.9%-109.9% |
| LLOQ | 0.25 ng/mL |
| ULOQ | 25 ng/mL |
| Detection limit | 0.0736 ng/mL |
| Repeatability | 3.4%-7.9% (CV≤20%) |
| Specificity | No cross-reactivity with commonly protein analogs (e.g. BSA, RNase A, DNase I), HCP (e.g. CHO, 293T, E.coli, Vero), and genomic DNA (e.g. Hansenula polymorpha, 293T, E.coli). |
| Robustness | Incubate at 25℃±5℃, CV≤ 20% |
| Instrument suitability not limited to Thermo Multiskan FC, Bio-Tek Synergy2 |
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