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Human TLR4(Toll-Like Receptor 4) ELISA Kit

Citations(7)Uniprot : O00206

Cat.No.:E-EL-H6123
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	31.25-2000 pg/mL
Sensitivity	18.75 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 < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human TLR4 concentrations in serum, plasma and other biological fluids.

Test Principle

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 TLR4. 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 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 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 TLR4. You can calculate the concentration of Human TLR4 in the samples by comparing the OD 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 ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 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), 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°C
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)	93.09	231.76	832.13	93.11	217.41	859.18
Standard deviation	5.17	10.52	36.11	4.96	10.39	42.01
CV (%)	5.55	4.54	4.34	5.33	4.78	4.89

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)	98-110	105
EDTA plasma (n=8)	91-105	99
Cell culture media (n=8)	92-103	95

Linearity

Samples were spiked with high concentrations of Human TLR4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: O00206
Synonyms	TLR-4, ARMD10, CD284, TOLL, hToll
Research Area	Cancer, Cardiovascular, Immunology, Microbiology

Assay Procedures

	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

Citations

Frontiers in Medicine (2022) IF: 5.058

Elevated serum TLR4 level as a potential marker for postsurgical chronic pain in pediatric patients with different approaches to analgesia.

DOI: 10.3389/fmed.2022.897533
	Sample: plasma

Nutrients (2021) IF: 5.719

Comparison of Simplified Creatinine Index and Systemic Inflammatory Markers for Nutritional Evaluation of Hemodialysis Patients

DOI: 10.3390/nu13061870
PMID: 34070850	Sample: Plasma

BRITISH JOURNAL OF NUTRITION (2022) IF: 4.125

Effects of inulin supplementation on inflammatory biomarkers and clinical symptoms of women with obesity and depression on a calorie-restricted diet: a randomized controlled clinical trial

DOI: 10.1017/S000711452200232X
PMID: 36059088	Sample: serum

STEM CELLS AND DEVELOPMENT (2015) IF: 3.727

Involvement of Immune Responses in the Efficacy of Cord Blood Cell Therapy for Cerebral Palsy

DOI: 10.1089/scd.2015.0074
	Sample: Serum

International Journal of Rheumatic Diseases (2022) IF: 2.558

The role of S100A12 and Toll-like receptor 4 in assessment of disease activity in familial Mediterranean fever and juvenile idiopathic arthritis

DOI: 10.1111/1756-185X.14475
PMID: 36302511	Sample: blood

ARCHIVES OF PHYSIOLOGY AND BIOCHEMISTRY (2017) IF: 1.22

Reduced soluble Toll-like receptors 2 in type 2 diabetes

DOI: 10.1080/13813455.2017.1401642
	Sample: Serum

EPILEPTIC DISORDERS (2020) IF: 1.276

HMGB‐1, TLR4, IL‐1R1, TNF‐α, and IL‐1β: novel epilepsy markers?

DOI: 10.1684/epd.2020.1155
	Sample: serum

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