Components:
HSA Calibration Standard
Anti-HSA Microtiter Strips
Diluent
Wash Buffer Concentrate (10×)
Anti-HSA:HRP (100×)
TMB Substrate
Stop Solution
Sealing Film
This kit is based on the solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect Deoxyribonuclease Ⅰ (DNase Ⅰ). A sheep polyclonal antibody specific to DNase Ⅰ was employed in the assay to capture any remaining DNase Ⅰ impurities in the sample. Both the calibration standards and test samples were simultaneously added to the microtiter plate coated with the affinity purified capture antibody, and followed by incubation and washing. The biotinylated antibody was added to the microtiter plate to bind the DNase Ⅰ and then reacted with streptavidin labeled 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 colored 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 DNase Ⅰ concentration in the calibration standard and the samples. The concentration of DNase Ⅰ in the samples can be calculated using a dose-response curve.
| Linearity & Range | 0.5-32 ng/mL, R2>0.990 | ||
| Accuracy | Recovery Rate= 89.2%-109.4% | ||
| LLOQ | 0.5 ng/mL | ||
| ULOQ | 32 ng/mL | ||
| Detection limit | 0.1 ng/mL | ||
| Precision | 6.6%-8.7% (CV≤20%) | ||
| Specificity | No cross-reactivity with Vero, HEK293T, CHO, E.coli, P.pastoris and SF9 HCP. | ||
| Robustness | Incubate at 25±3℃, CV≤ 20% | ||
| Instrument suitability not limited to MD Spectra Max ABS and Thermo Multiskan FC | |||
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