BioReagent,for enzyme immunoassay(ELISA) BioReagent,per il test immunoenzimatico (ELISA) for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
This kit adopts the double-antibody sandwich ELISA method to detect Benzonase content in samples. Benzonase-specific antibodies are pre-coated on microplate wells. During detection, standards and samples are added separately into antibody-coated wells. Benzonase in samples binds to the immobilized antibodies. After incubation, unbound substances are washed away. HRP-labeled Benzonase detection antibody is added to form antibody-antigen-HRP antibody complexes. TMB substrate is added for color development. The color intensity is proportional to Benzonase concentration. The residual Benzonase in samples is determined by detecting color development of Benzonase standards.
Detection Range: 0.094~6 ng/mL
Quantitation Limit: 0.094 ng/mL
Limit of Detection: 0.019 ng/mL
Required Instruments
Microplate reader, microplate constant temperature shaker, vortex mixer, pipettes, plate sealing film, etc.
Reagent Preparation
1. Equilibration: Allow all required reagents to equilibrate at room temperature (25±3℃) for at least 30 minutes. Determine the number of wells needed, take out required microplate strips, and return unused strips to aluminum foil bags with desiccant. Reseal and store at 2–8℃.
2. Prepare 1× Washing Buffer: Dilute 20× Washing Buffer with deionized water at a volume ratio of 1:20 according to required volume, mix well for standby use.
3. Standard Solution Dilution (Example):
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Assay Procedure
1. Mix all reagents equilibrated to room temperature gently before use.
2. Pipette 100 μL of standard solutions (St1~St8) or test samples into corresponding wells. Seal the plate with a plate sealer, then incubate at 25±3 ℃ and 400–500 rpm for 1 hour.
3. Washing: Discard the liquid in wells. Add at least 350 μL of 1× Washing Buffer to each well (using a multichannel pipette or wash bottle). Discard residual liquid and pat the plate dry on lint-free absorbent paper. Repeat washing for 3 cycles.
(Note: Do not allow wells to dry excessively for a long time before adding Benzonase detection antibody. Improper washing may cause high CV between duplicate wells and abnormal absorbance signals.)
4. Add 100 μL of HRP-labeled Benzonase detection antibody to each well. Seal the plate and incubate at 25±3 ℃, 400–500 rpm for 30 minutes.
5. Washing: Discard the liquid in wells. Add at least 350 μL of 1× Washing Buffer to each well. Discard liquid and pat the plate dry. Repeat washing 3 times.
(Note: Avoid prolonged excessive drying of wells prior to TMB substrate addition.)
6. Color development: Add 100 μL TMB substrate to each well, and incubate for 20 minutes at room temperature in the dark.
7. Termination: Add 100 μL Stop Solution to each well, tap the plate gently to mix thoroughly and terminate the reaction.
8. Detection: Measure the absorbance at 450 nm within 10 minutes after adding Stop Solution.
Data Calculation
It is recommended to perform logarithmic transformation of concentration and fit standard curve with 4-Parameter Logistic regression model.
Table 1 Typical Standard Curve Data
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Typical Standard Curve (4PL Logistic Model)
Performance
1. Limit of Detection (LOD): 0.019 ng/mL. Calculated by substituting the average absorbance of 20 blank controls plus 2 standard deviations into the standard curve.
2. Limit of Quantitation (LOQ): 0.094 ng/mL. The lowest concentration with coefficient of variation (n=6) ≤ 20%.
3. Linearity: Standard curves are plotted with serially diluted standard solutions, with R² ≥ 0.98.
4. Repeatability: Evaluated by detecting three samples of known concentrations for 20 replicates.
5. Intermediate Precision: Evaluated by detecting three samples of known concentrations 4 times by different operators on different days.
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6. Recovery: The recovery rates of spiked Benzonase at different levels within the full detection range in biological matrix were verified.
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Precautions
1. For research use only, not for clinical diagnosis or treatment.
2. Store reagents as labeled. Equilibrate reagents and samples to room temperature before detection.
3. Equilibrate pre-coated microplates to room temperature before opening packages. Return unused strips to sealed bags and store at 2–8℃. Seal all leftover reagents properly.
4. Mix Benzonase standard and HRP detection antibody thoroughly before use.
5. Pat washed wells dry on lint-free paper. Do not over-dry wells, which will increase CV and blank OD deviation.
6. TMB substrate should be colorless and transparent. Pale blue TMB indicates contamination and cannot be used.
7. Samples with extreme pH (<5.0 or >8.5), high salt, polysaccharide, urea, organic solvent or detergent may result in low recovery.
8. Due to complex biological matrix, use positive controls and duplicate wells to ensure accurate results.
Safety Instructions
1. Stop Solution is acidic. Operate with caution.
2. Wear proper personal protective equipment: lab coat, gloves, mask, goggles, etc.
| Item No. | Appearance | Components | 96 T | Storage |
| B1522792A | Liquid | Benzonase Standard | 1000 ng/ml, 0.8 ml, 1 Vial | 2-8℃ |
| B1522792B | Liquid | Benzonase Detection Antibody (HRP labeled) | 12 ml, 1 Vial | 2-8℃. Store in the dark. |
| B1522792C | Liquid | TMB Substrate | 12 ml, 1 Vial | 2-8℃. Store in the dark. |
| B1522792D | Liquid | Stop Solution | 12 ml, 1 Vial | 2-8℃ |
| B1522792E | Liquid | Sample Diluent | 50 ml, 1 Vial | 2-8℃ |
| B1522792F | Liquid | Wash Buffer (20×) | 50 ml, 1 Vial | 2-8℃ |
| B1522792G | — | Pre-coated Assay Plate | 96 wells，1 Plate | 2-8℃ |
Comprehensive hazard, handling, storage, and regulatory compliance document.
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