Product description

Salt Active UltraNuclease is a non-specific endonuclease which derived from Marine microorganisms and is expressed and purified in Escherichia coli (E. coli) by genetically engineered. This nonspecific, recombinant endonuclease has optimum activity at high salt concentration (500mM NaCl), which can improve efficiency and yield in various workflows.

This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residual Salt Active UltraNuclease in samples. First, add the Salt Active UltraNuclease standard and test sample to the Anti-Salt Active UltraNuclease coated microtiter strips (36719-A), then add the diluted biotin labeled Anti-UltraNuclease antibody (36719-C), and finally add Streptavidin-HRP (36719-D) to form an antibody + antigen + antibody-Biotin + SA-HRP complex. Subsequently, add TMB substrate (36719-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 Salt Active UltraNuclease in the sample.

The detection range of this kit is 0.047-3 ng/mL; the lower detection limit is 0.024 ng/mL.

Feature

 

• Highly sensitive: Detecting as little as 0.024 ng/mL of Salt Active UltraNuclease residuals in in-process and final biological samples.
• Ensures accuracy: The Salt Active UltraNuclease residuals can be accurately detected with a recovery of 75%-125%.
• Specificity: Cooperate use with YEASEN Salt Active UltraNuclease 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 Salt Active UltraNuclease test in biological products

Specification

Sensitivity	0.024 ng/mL (range 0.047~3 ng/mL)
Assay Time	<4 hours
Assay Principle	Two-site immunoenzymetric assay
Signal Amplification	Biotin-Streptavidin system
Detection Wavelength	450nm-630nm

Components

Components No.	Name	36719ES48	36719ES96
36719-A	Anti-Salt Active UltraNuclease coated microtiter strips	48 T	96 T
36719-B*	Standard: Salt Active UltraNuclease	1 vial	2 vial
36719-C	Detection Antibody：Biotin-conjugated Antibodies	60 μL	120 μL
36719-D	Streptavidin-HRP	30 μL	60 μL
36719-E	Dilution Buffer 1	25 mL	50 mL
36719-F	Wash Buffer Concentrate (20×)	25 mL	50 mL
36719-G	Dilution Buffer 2	15 mL	30 mL
36719-H	TMB Substrate	8 mL	15 mL
36719-I	Plate Sealer	3 each	5 each

* 1. The stop solution is not provided in this kit, it need to be prepared by user. 1M HCl can be used as the stop solution;

* 2. The Standard (36719-B) is Lyophilized powder.

Storage

This product should 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.

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

 Figures

• Specificity Demonstration

Figures 1. The specificity of Salt Active UltraNuclease ELISA kit was shown in the figure above

7 samples were estimated for the non-specific reaction with the antibodies in the kit. The concentration of the 7 samples were all below the QL (0.047ng/mL).

• Precision Display

A）Repeatability

Spiked sample	Intra-Assay
	Duplicates	Average Conc. (ng/mL)	CV (%)
S-1.5 ng/mL	10	1.344	2.97%
S-0.375 ng/mL	10	0.260	2.27%
S-0.094 ng/mL	10	0.061	2.56%

B）Intermediate precision

Spiked sample	Inter-Assay
	Duplicates	Average Conc. (ng/mL)	CV (%)
S-1.5 ng/mL	30	1.335	5.70%
S-0.375 ng/mL	30	0.258	5.59%
S-0.094 ng/mL	30	0.064	6.82%

Figures 2. The precision of Salt Active UltraNuclease 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 5% (Figure 2A). 3 lot of Salt Active UltraNuclease ELISA kits were tested and the CV% were lower than 10% (Figure 2B).

• Accuracy Validation

Dilution factor	Dilution recovery (%)	Range of Dilution recovery（%）
1:2	92.5	87.5-98.9
1:4	87.0	79.0-95.2
1:8	92.6	78.9-106.2
1:16	96.9	79.1-118.1
Spiked samples	Spike recovery (%)	Range of Spike recovery（%）
S-0.75ng/mL	105.3	91.0-122.1
S-0.18ng/mL	96.6	86.5-105.5

Figures 3. The accuracy of UCF.ME™ UltraNuclease ELISA was assayed as the following scheme

The protein sample with high Salt Active UltraNuclease concentration was serial 2 fold diluted and also spiked with 0.75ng/mL and 0.18ng/mL Salt Active UltraNuclease. The dilution recovery and the spike recovery were all within 75%-125% (Figures 3).