Human NLRP3 ELISA Kit

Product Name:	Human NLRP3 ELISA Kit
Product Code:	AEES00672
Assay Type:	Sandwich
Format:	96T
Assay Time:	3.5h
Reactivity:	Human
Detection Range:	0.16-10 ng/mL
Sensitivity:	0.09 ng/mL
Sample Volume:	100μL
Specificity:	This kit recognizes Human NLRP3 in samples. No significant cross-reactivity or interference between Human NLRP3 and analogues was observed.
Reproducibility:	Both intra-CV and inter-CV are < 10%.
Application:	This ELISA kit applies to the in vitro quantitative determination of Human NLRP3 concentrations in.

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human NLRP3. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human NLRP3 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human NLRP3, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human NLRP3. You can calculate the concentration of Human NLRP3 in the samples by comparing the OD of the samples to the standard curve.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NLRP3 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NLRP3 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	0.47	0.84	4.71	0.46	0.78	4.71
Standard deviation	0.03	0.04	0.17	0.03	0.04	0.16
CV (%)	6.38	4.76	3.61	6.52	5.13	3.40

Recovery

The recovery of Human NLRP3 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	85-96	91
EDTA plasma (n=8)	87-98	92
Cell culture media (n=8)	92-106	98

Kit Components:

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	8 wells x 12 strips	-20℃, 6 months
Reference Standard	2 vials
Concentrated Biotinylated Detection Ab (100×)	1 vial, 120 μL
Concentrated HRP Conjugate (100×)	1 vial, 120 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

1.	Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
2.	Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
3.	Aspirate and wash the plate for 3 times
4.	Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
5.	Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
6.	Add 50μL Stop Solution
7.	Read the plate at 450nm immediately. Calculation of the results