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Rat TLR4(Toll-Like Receptor 4) ELISA Kit

Cat.No.:E-EL-R0990
Reactivity: Rat

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat TLR4 in samples. No significant cross-reactivity or interference between Rat 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 Rat 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 Rat 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 Rat 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 Rat 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 Rat TLR4. You can calculate the concentration of Rat 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 Rat TLR4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat 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 (ng/mL)	0.95	2.18	8.78	0.91	2.03	8.85
Standard deviation	0.06	0.13	0.29	0.05	0.11	0.31
CV (%)	6.32	5.96	3.30	5.49	5.42	3.50

Recovery

The recovery of Rat 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)	93-111	101
EDTA plasma (n=8)	94-108	101
Cell culture media (n=8)	92-109	100

Linearity

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

Target Information

Synonyms	TLR-4, ARMD10, CD284, TOLL
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

Food & Function (2021) IF: 5.396

Dry heated sorghum BRS 305 hybrid flour as a source of resistant starch and tannins improves inflammation and oxidative stress in Wistar rats fed with a high-fat high-fructose diet

DOI: 10.1039/D1FO00802A
PMID: 34369542	Sample: liver

Food & Function (2022) IF: 5.396

Black corn (Zea mays L.) whole flour improved the antioxidant capacity and prevented the adipogenesis in mice fed a high-fat diet.

DOI: 10.1039/D1FO04205J
	Sample: adipose tissue

Frontiers in Pharmacology (2021) IF: 5.811

Rivaroxaban Modulates TLR4/Myd88/NF-Kβ Signaling Pathway in a Dose-Dependent Manner With Suppression of Oxidative Stress and Inflammation in an Experimental Model of Depression

DOI: 10.3389/fphar.2021.715354
PMID: 34630092	Sample: hippocampal tissue

Foods (2022) IF: 4.35

Chia (Salvia hispanica L.) Flour and Oil Ameliorate Metabolic Disorders in the Liver of Rats Fed a High-Fat and High Fructose Diet

DOI: 10.3390/foods11030285
	Sample: liver tissue

BIOMEDICINE & PHARMACOTHERAPY (2020) IF: 4.545

Diacerein protects rats with liver ischemia/reperfusion damage: Down-regulation of TLR4/ NFκ-B signaling pathway

DOI: 10.1016/j.biopha.2020.111063
PMID: 33348310	Sample: Tissue homogenate

Journal of Functional Foods (2020) IF: 3.701

Regulation of inflammatory response and oxidative stress by tocotrienol in a rat model of non-alcoholic fatty liver disease

DOI: 10.1016/j.jff.2020.104209
	Sample: serum and liver

MEDIATORS OF INFLAMMATION (2020) IF: 3.758

Paeonol Attenuates Methotrexate-Induced Cardiac Toxicity in Rats by Inhibiting Oxidative Stress and Suppressing TLR4-Induced NF-κB Inflammatory Pathway

DOI: 10.1155/2020/8641026
	Sample: Soil,Water,Oil,Plant Tissue,Plant Fluid,Antibody,F

JOURNAL OF MOLECULAR NEUROSCIENCE (2020) IF: 2.678

Neuroprotective Effects of Ethyl Pyruvate against Aluminum Chloride-Induced Alzheimer’s Disease in Rats via Inhibiting Toll-Like Receptor 4

DOI: 10.1007/s12031-020-01489-9
	Sample: Tissue homogenate

ANAEROBE (2022) IF: 2.837

The comparison of changes in fecal and mucosal microbiome in metabolic endotoxemia induced by a high-fat diet

DOI: 10.1016/j.anaerobe.2022.102615
PMID: 35850456	Sample: serum,colon

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