Product description

T7 RNA polymerase that synthesizes RNA complementary to reverse single-stranded DNA downstream of the promoter using double-stranded DNA containing the T7 promoter sequence (5'-TAATACGACT CACTATAG*-3') as a template and NTP as a substrate.

This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residual T7 RNA polymerase in samples. First, add the T7 RNA polymerase standard and test sample to the Anti-T7 RNA Polymerase coated microtiter strips (36705-A), then add the diluted biotin labeled Anti-T7 RNA polymerase antibody (36705-C), and finally add Streptavidin-HRP (36705-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, add TMB substrate (36705-H) into the complex to observe color reaction after washing the complex. TMB is converted into blue under the catalysis of HRP enzyme and finally converted into yellow in the presence of acid, and the shade of color is positively correlated with the amount of T7 RNA polymerase in the sample.

The detection range of this kit is 1~32 ng/mL; the lower detection limit is 0.318 ng/mL.

Feature

• Highly sensitive: Detecting as little as 0.318 ng/mL of T7 RNA Polymerase residuals in in-process and final biological samples.
• Ensures accuracy: The T7 RNA Polymerase residuals can be accurately detected with a recovery of 75%-125%.
• Specificity: Cooperate use with YEASEN T7 RNA Polymerase and can accurately detect the residuals.
• Stable: The difference between lot-to-lot is low, the kit property is not impacted under 37℃ for 7 days.
• Easy signal collection: Using Biotin-Streptavidin system, the signal can be stably enlarged.

Application

Residual T7 RNA Polymerase test in biological products

Specification

Components

^*The Standard (36705-B) is Lyophilized powder.

Storage

This product should be stored at 2°C ~8°C. Unopened product is valid for one year.

^*Upon receipt of the kit, please check whether all components are complete and immediately store them in corresponding condition.

Figures

Table 1. Linearity Range: 1～32 ng/mL ，R2=0.999 ，CV≤5%。

• Specificity Demonstration

Figure 1. The specificity of T7 RNA Polymerase ELISA kit was shown in the figure above

8 samples were estimated for the non-specific reaction with the antibodies in the kit. The concentration of the 8 samples were all below the Limit of Quantitation (LOQ) (0.318ng/mL).

• Precision Display

A) Repeatability

B) Intermediate precision

Table 1. The precision of T7 RNA Polymerase ELISA was assayed as the above scheme

10 duplicated were tested at 3 concentrations on the standard curve, and all the CV% were lower than 10% (Table 1A). 3 lot of Salt Active UltraNuclease ELISA kits were tested and the CV% were lower than 15% (Table 1B).

l Accuracy Validation

Three concentrations of T7 standards, high, medium and low, were added to the mRNA purified samples in equal proportions, while the high value spiked samples were made multiplicative dilutions.

Spiked recovery = measured value / (spiked standard x 50% + spiked specimen value x 50%)

Table 2. Accuracy Validation

The linear and spiked recoveries of T7 in the doubly diluted samples, as well as the samples with different spiked concentrations, and in the unspiked samples, ranged from 80% to 120% (Table 2). 

Documents:

Safety Data Sheet

Manuals