Human TLR4 (Toll-Like Receptor 4) CLIA Kit

SKU: E-CL-H0986

$ 579⁹⁵

Human TLR4 (Toll-Like Receptor 4) CLIA Kit

Detection Range	62.50-4000 pg/mL
Sensitivity	37.50 pg/mL

Product Details

Properties

Assay type	Sandwich-CLIA
Format	96T/48T
Assay time	3.5h
Detection range	62.50-4000 pg/mL
Sensitivity	37.50 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human TLR4 in samples. No significant cross-reactivity or interference between Human TLR4 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 15%.
Application	This CLIA kit applies to the in vitro quantitative determination of Human TLR4 concentrations in serum, plasma and other biological fluids.

Test Principle

This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human TLR4. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TLR4 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 TLR4, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human TLR4. You can calculate the concentration of Human TLR4 in the samples by comparing the RLU value of the samples to the standard curve.

Kit Components & Storage

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 CLIA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 12 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 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 A	1 vial, 5 mL	2-8℃(Protect from light)
Substrate Reagent B	1 vial, 5 mL	2-8℃(Protect from light)
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TLR4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TLR4 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 (pg/mL)	193.73	335.07	1584.16	207.30	367.47	1733.16
Standard deviation	20.79	30.79	127.52	21.25	40.16	191.86
CV (%)	10.73	9.19	8.05	10.25	10.93	11.07

Recovery

The recovery of Human TLR4 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)	85-97	91
EDTA plasma(n=8)	101-116	107
Cell culture media(n=8)	99-112	105

Target Information

Synonyms	TLR-4, ARMD10, CD284, TOLL, hToll
Research Area	Cancer, Cardiovascular, Immunology, Microbiology

Assay Procedures

	1.Add 100 μL of standard or sample to each well. Incubate for 90 min at 37℃.
	2.Remove the liquid.
	3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times.
	4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃. Aspirate and wash 5 times.
	5.Add 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃.
	6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results.