AffiELISA® RNase Inhibitor ELISA Kit
AffiELISA® RNase Inhibitor ELISA Kit is a high-sensitivity immunoassay designed for the quantitative detection of RNase inhibitor in biological or manufacturing samples.
This assay is especially suited for monitoring residual RNase inhibitor during purification processes to confirm product purity and integrity. With high specificity and reproducibility, the kit supports critical research and manufacturing needs, particularly where RNA stability is essential, such as in vaccine development, gene therapy, and nucleic acid-based therapeutics.
Intended Use:
AffiELISA® RNase Inhibitor ELISA Kit is intended for the quantitative detection of RNase inhibitor in research and bioprocessing samples.
Kit components:
| Components | Description |
| Pre-coated Microplate | Wells coated with a capture antibody specific to the target analyte. |
| Standards | A set of solutions containing known concentrations of the target analyte for creating a standard curve. |
| Detection Antibody (1X) | A solution containing a primary antibody that specifically binds to the captured target analyte. |
| HRP-Streptavidin (1X) | A solution of horseradish peroxidase (HRP) conjugated to streptavidin, used for signal amplification (assuming the detection antibody is biotinylated). |
| Wash Buffer | A solution used to remove unbound substances during washing steps. |
| Substrate Solution | A solution that reacts with the HRP enzyme to produce a colorimetric signal. |
| Stop Solution | A solution used to halt the enzymatic reaction and stabilize the color. |
Materials Required But Not Supplied:
- Microplate reader capable of reading absorbance at D450/650 nm.
- Microplate shaker.
Procedure workflow:
1. Prepare Reagents:
- Assemble all necessary reagents, including standards, samples, wash buffer, detection antibody, HRP-streptavidin, and substrate solution.
2. Add Standards and Samples:
- Pipette 100 µL of standards and samples into the designated wells of the microplate.
3. First Incubation:
- Incubate the plate at room temperature (25°C) with shaking at 500 rpm for 60 minutes.
4. First Wash:
- Discard well contents.
- Blot plate dry on absorbent paper.
- Add 300 µL wash buffer.
- Incubate 15-30 seconds.
- Discard wash buffer.
- Blot plate dry.
- Repeat wash cycle 3 times.
5. Add Detection Antibody:
- Pipette 100 µL of 1X detection antibody into each well.
6. Second Incubation:
- Incubate the plate at room temperature (25°C) with shaking at 500 rpm for 60 minutes.
7. Add HRP-Streptavidin:
- Pipette 100 µL of 1X HRP-streptavidin into each well.
8. Third Incubation:
- Incubate the plate at room temperature (25°C) with shaking at 500 rpm for 30 minutes.
9. Second Wash: (Same as First Wash)
- Discard well contents.
- Blot plate dry on absorbent paper.
- Add 300 µL wash buffer.
- Incubate 15-30 seconds.
- Discard wash buffer.
- Blot plate dry.
- Repeat wash cycle 3 times.
10. Add Substrate Solution:
- Pipette 100 µL of substrate solution into each well.
11. Incubation:
- Incubate the plate at 37°C in the dark for 10 minutes.
12. Add Stop Solution:
- Pipette 50 µL of stop solution into each well.
13. Read Absorbance:
- Measure the absorbance of each well at OD450/650 nm using a microplate reader.