Description
Double-stranded RNA (dsRNA) emerges as a by-product during the in vitro transcription of mRNA. This by-product possesses immunogenic properties within the human body, capable of provoking an immune response, consequently diminishing mRNA levels. This renders dsRNA a troublesome process impurity necessitating thorough elimination and stringent control of its residual presence.
To address this concern, the Double-stranded RNA (dsRNA) ELISA detection kit employs the experimental principles of a double-antibody sandwich enzyme-linked immunoassay (ELISA) for the quantification of residual dsRNA. The Kit can not only be used in the detection of regular dsRNAs, but also in detection of peudo UTP, N1-Me-peudo UTP and 5-OMe-UTP modified dsRNAs.
The procedure involves adding both a standard and the test sample onto an enzyme plate (designated as 36717-A) previously coated with anti-dsRNA antibodies. Subsequently, biotin-labeled dsRNA detection antibodies (36717-F) are added in a diluted form. Finally, Streptavidin-HRP (SA-HRP) (36717-G) is introduced to create a complex comprising antibodies, antigens, biotin, and SA-HRP.
Following plate washing, the addition of TMB chromogenic solution (36717-K) initiates color development. Under the influence of HRP enzyme catalysis, TMB undergoes a transformation from colorless to blue, which is then halted through the application of the stop solution (36717-L). Ultimately, this transformation results in a yellow hue, the intensity of which correlates positively with the quantity of dsRNA identified in the sample.
This kit serves a versatile purpose, enabling optimization of the biological product purification process, impurity control during intermediate stages, and release testing for final product assessment.
Components
|
No. |
Name |
36717ES48 |
36717ES96 |
|
36717-A |
Anti-dsRNA coated microtiter strips |
48 T |
96 T |
|
36717-B |
STD 1: unmodified, 5 ng/μL |
7.5 μL |
15 μL |
|
36717-C |
STD 2: pUTP modified, 5 ng/μL |
7.5 μL |
15 μL |
|
36717-D |
STD 3: N1-Me-pUTP modified, 5 ng/μL |
7.5 μL |
15 μL |
|
36717-E |
STD 4: 5-OMe-UTP modified, 5 ng/μL |
7.5 μL |
15 μL |
|
36717-F |
Detection Antibody：Biotin-conjugated Antibodies (100x) |
60 μL |
120 μL |
|
36717-G |
Streptavidin-HRP (100x) |
60 μL |
120 μL |
|
36717-H |
Dilution Buffer 1 |
25 mL |
50 mL |
|
36717-I |
Dilution Buffer 2 |
15 mL |
30 mL |
|
36717-J |
Wash Buffer Concentrate（20×) |
20 mL |
40 mL |
|
36717-K |
TMB Substrate |
6 mL |
12 mL |
|
36717-L |
Stop Solution |
3 mL |
6 mL |
|
36717-M |
Plate Sealer |
2 |
4 |
Shipping and Storage
The kit is shipped with an ice pack and can be stored at 2°C ~8°C. Unopened product is valid for one year. Once the reagent is opened, it is valid for half a year. Never freeze this product.