RNase Inhibitor ELISA Kit,Absin_specification/price/image

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Product Specification

Usage

1. Preparation:
Before using all components of reagent and sample under test needs to be restored at room temperature.
1 and 1 x for Wash Buffer preparation: 20 x will Wash Buffer balance to room temperature to crystallization completely dissolved, using deionized water or ultrapure water diluted 20 times after blending, diluted 1 x for Wash Buffer every hole are use 3 ml, in 2-8 ℃ can keep stable for 1 week.
2, RNase Inhibitor Detection Antibody Dilution: before using blender, and then use Dilution Buffer 2 will RNase Inhibitor DetectionAntibody according to 1: 100 dilution ratio of dilution.
3. Preparation of the substrate solution: Color Reagent A and Color Reagent B were mixed according to 1:1, and then mixed for immediate use. The dosage of each hole 100 mu L. Do not use if the mixed substrate solution turns blue. 4. RNase Inhibitor dilution of Standard: 6μL RNase Inhibitor Standard stock solution (10μg/mL) was diluted 100-fold with 594 μl dilution Buffer 1 as the first gradient (100ng/mL). Then take 100 ng/mL concentration sample 300 mu L to new EP tube, add 300 mu L Dilution Buffer1, double gradient Dilution, Dilution into 50, 25, 12.5, 6.25, 3.125 and 1.5625 ng/mL, a total of seven levels.

In order to ensure the validity of the experimental results, please use the newly prepared standard solution for each experiment.

2, the operation process:
All operating at room temperature, it is recommended that the standard and the sample under test to determine double complex hole.
1, sample incubation: will be diluted RNase Inhibitor of Standard and sample under test in enzyme label plate, respectively, 100 mu L/hole, blending, cover the seal plate membrane, 37 ℃ for 2 h incubation.
2, washing board: abandon to liquid in the hole, in 1 x for Wash Wash Buffer board, 300 mu L/hole, soak 30 s, washing board 5 times, the last time to pat dry residual liquid.
3, Inhibitor Detection Antibody incubation: after will dilute the RNase Inhibitor Detection Antibody in enzyme label plate, 100 mu L/hole, blending, cover the seal plate membrane, 37 ℃ for 1 h incubation.
4. Wash the plate: discard the liquid in the hole, wash the plate with 1×Wash Buffer, 300μL/ well, soak for 30s, wash the plate 5 times, and pat the residual liquid dry as much as possible for the last time.
5. Color development: The prepared substrate solution was added to the microplate, 100μL/ well, mixed, covered with plate sealing membrane, and incubated at 37 ° C in the dark for 20min.
6. Termination: Add Stop Reagent to the microplate, 50μL/ well, and gently shake the microplate until the color is uniform to ensure that the color development time of each well is the same.
7, reading value: enzyme label plate to be included in the enzyme standard instrument, read in 450 nm wavelength absorbance, read values within 20 minutes.

3. Data processing
If the OD value of the sample to be tested exceeded the OD value of the highest point of the standard curve, the sample should be diluted and measured again.
With standard concentration as the abscissa, standard values for the vertical suction light drawing standard curve. ELISACalc.exe regression and fitting calculation software is recommended for four-parameter fitting to calculate the concentration of RNase Inhibitor in samples.

Theory

The kit is a double-antibody sandwich ELISA method, which is precoated with RNase Inhibitor to capture antibody. After adding the sample, the sample is captured to form antibody-antigen complex, and then HRP-labeled RNase Inhibitor is added to detect antibody. An antibody-antigen-antibody "sandwich" complex was formed. Finally, TMB was added for color development, and the absorbance (OD) value was read at 450nm wavelength after the reaction was terminated. The RNase Inhibitor content in the sample was positively correlated with the OD value.

Description

RNase Inhibitor can specifically inhibit the activities of RNase A, B and C, and can form a 1:1 complex with RNase, thereby inhibiting its activity.
In the process of drug mRNA vaccine production, RNase Inhibitor can effectively inhibit the reaction system of possible RNase activity, so as to protect the mRNA degradation. According to the regulation, need to concentrate to the residue detection of mRNA vaccine, including RNase Inhibitor as protein product is belong to the category of protein residue test items, so we need the residues were quantitative detection.
This kit, double antibody sandwich enzyme-linked immunoassay (ELISA) to detect RNase Inhibitor, with high sensitivity and specificity of detection and quantitative analysis of RNase Inhibitor content.

Product Features:
Sensitivity: 3.125 ng/ml
Detection Range: 3.125 100 ng/ml

Composition

Components	Descriptions	Size
RNase Inhibitor Microplate	Package is capture antibody	96T/Kit
RNase Inhibitor Detection Antibody	HRP was labeled and the dilution ratio was 1:100	150μl
RNase Inhibitor Standard	Restructuring RNase inhibitor (5 mu g/ml)	150μl
20×Wash Buffer	scrubbing solution	30ml×2
Sample Dilution Buffer	Sample diluent	30ml
Conjugate Dilution Buffer	enzyme diluent	15ml
Color Reagent A	Color liquid A	6ml
Color Reagent B	Color liquid B	6ml
Stop Reagent	stop buffer	6ml
Sealing Film	sealing film	4

General Notes

1, before the experiment should be in accordance with the requirements of relevant experimental reagent balance to room temperature.
2, each reagent should be fully mixed before use to ensure the uniformity and accuracy of the reagent.
3, strictly control the reaction temperature and reaction time.
4, measurements can not beyond the normal range.
5, the number of washing plate, the amount of washing plate liquid, the soaking time of washing plate liquid, and the strength of washing plate are the problems that should be paid attention to.
6, when the termination reaction to avoid microporous of bubbles.
7. The corresponding standard curve should be prepared for each test, and the standard curve of different kits and different days should not be mixed.
8, pay attention to the change in time between different samples and steps with tray and spear, avoid cross contamination.
9, reading the light absorption value should be completed within 20min after adding the termination solution.
10, color with high concentration may produce black floc, belongs to the normal phenomenon, lightly don't affect the reading result.

Storage Temp.

Dry and keep away from light at 2-8 ° C, valid for 12 months.

Test Range

3.125-100 ng/ml

AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.