Human toll-like receptor 4,TLR4 ELISA Kit

Product Details

Citations

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Target Background

Product Details

Target Name

toll-like receptor 4

Alternative Names

ARMD10 ELISA Kit; CD284 ELISA Kit; CD284 antigen ELISA Kit; ESOP 1 ELISA Kit; ESOP1 ELISA Kit; Homolog of Drosophila toll ELISA Kit; hToll ELISA Kit; LY 96 ELISA Kit; LY96 ELISA Kit; Lymphocyte antigen 96 ELISA Kit; md 2 ELISA Kit; MD 2 protein ELISA Kit; MD2 protein ELISA Kit; Myeloid differentiation protein 2 ELISA Kit; Protein MD 2 ELISA Kit; Protein MD2 ELISA Kit; TLR 4 ELISA Kit; TLR4 ELISA Kit; TLR4_HUMAN ELISA Kit; TOLL ELISA Kit; Toll like receptor 4 ELISA Kit; Toll-like receptor 4 ELISA Kit

Abbreviation

TLR4

Uniprot No.

O00206

Species

Homo sapiens (Human)

Sample Types

serum, plasma, tissue homogenates

Detection Range

0.156 ng/mL-10 ng/mL

Sensitivity

0.039 ng/mL

Assay Time

1-5h

Sample Volume

50-100ul

Detection Wavelength

450 nm

Research Area

Immunology

Assay Principle

quantitative

Measurement

Sandwich

Precision

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of human TLR4 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
	Sample	Serum(n=4)
1:5	Average %	85
	Range %	80-93
1:10	Average %	95
	Range %	89-100
1:20	Average %	97
	Range %	93-104
1:40	Average %	95
	Range %	91-99

Recovery

The recovery of human TLR4 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.

Sample Type	Average % Recovery	Range
Serum (n=5)	97	89-103
EDTA plasma (n=4)	97	90-102

Typical Data

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

ng/ml	OD1	OD2	Average	Corrected
10	2.463	2.547	2.505	2.372
5	2.121	2.172	2.147	2.014
2.5	1.540	1.516	1.528	1.395
1.25	0.725	0.747	0.736	0.603
0.625	0.418	0.436	0.427	0.294
0.312	0.236	0.241	0.239	0.106
0.156	0.199	0.206	0.203	0.070
0	0.132	0.134	0.133

Materials provided

A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human TLR4 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
One vial Biotin-labeled TLR4 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
An instruction manual

Materials not provided

A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
An incubator can provide stable incubation conditions up to 37°C±5°C.
Centrifuge
Vortex
Squirt bottle, manifold dispenser, or automated microplate washer
Absorbent paper for blotting the microtiter plate
50-300ul multi-channel micropipette
Pipette tips
Single-channel micropipette with different ranges
100ml and 500ml graduated cylinders
Deionized or distilled water
Timer
Test tubes for dilution

ELISA Data Analysis

ELISA Standard Curve & Curve Expert software

Troubleshooting
and FAQs

ELISA kit FAQs

Storage

Store at 2-8°C. Please refer to protocol.

Lead Time

3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx

Description

This Human TLR4 ELISA Kit was designed for the quantitative measurement of Human TLR4 protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 0.156 ng/mL-10 ng/mL and the sensitivity is 0.039 ng/mL.

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Target Background

Function
(From Uniprot)

Cooperates with LY96 and CD14 to mediate the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MYD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Also involved in LPS-independent inflammatory responses triggered by free fatty acids, such as palmitate, and Ni(2+). Responses triggered by Ni(2+) require non-conserved histidines and are, therefore, species-specific. Both M.tuberculosis HSP70 (dnaK) and HSP65 (groEL-2) act via this protein to stimulate NF-kappa-B expression. In complex with TLR6, promotes sterile inflammation in monocytes/macrophages in response to oxidized low-density lipoprotein (oxLDL) or amyloid-beta 42. In this context, the initial signal is provided by oxLDL- or amyloid-beta 42-binding to CD36. This event induces the formation of a heterodimer of TLR4 and TLR6, which is rapidly internalized and triggers inflammatory response, leading to the NF-kappa-B-dependent production of CXCL1, CXCL2 and CCL9 cytokines, via MYD88 signaling pathway, and CCL5 cytokine, via TICAM1 signaling pathway, as well as IL1B secretion. Binds electronegative LDL (LDL(-)) and mediates the cytokine release induced by LDL(-). Stimulation of monocytes in vitro with M.tuberculosis PstS1 induces p38 MAPK and ERK1/2 activation primarily via TLR2, but also partially via this receptor. Activated by the signaling pathway regulator NMI which acts as damage-associated molecular patterns (DAMPs) in response to cell injury or pathogen invasion, therefore promoting nuclear factor NF-kappa-B activation.

Gene References into Functions

the presence of polymorphism Asp299Gly gene TLR4 in patients co-infected with HIV/HCV indicates a high risk of metabolic disturbances PMID: 30204113
high expression of (TLR4) is associated with Type 2 diabetes mellitus. PMID: 30403590
Data suggest that the toll-like receptor 4 (TLR4) mutant-specific conformational alterations may help in deciphering the mechanism of loss-of-function mutations. PMID: 28272553
TLR4 and TLR9 mRNA were elevated in blood samples from celiac disease patients compared to the healthy controls PMID: 30057921
high TLR4 expression level during acute rejection was associated with adverse kidney allograft outcome PMID: 29475090
the results of the present study showed significantly higher mRNA expression levels for TLR4 180days post-transplantation in the graft dysfunction group compared to well functioning graft group PMID: 29452169
the expression levels of TLR4/MyD88 were positively correlated with the metastatic potential of breast cancer cells and tumors. The expression levels of TLR4/MyD88 may be used as a biomarker to evaluate the prognosis and guide the treatment of patients with breast cancer. PMID: 30066873
These results suggest that celastrol exerts its protective effect partly via inhibiting the TLR4mediated immune and inflammatory response in steatotic HepG2 cells. PMID: 30015859
No association between the SNPs rs10983755 A/G, rs4986791 C/T, rs4986790 A/G, rs10759932 C/T, rs1927911 C/T, rs11536889 C/G, and rs12377632 C/T and heart transplant rejection was found PMID: 30177119
The results revealed that TLR4 and COX-2 were upregulated in PCa tissues; silencing of TLR4 or COX-2 inhibited PCa cell proliferation, migration, and invasion. PMID: 30098292
The role of IDO1-IDO2-AHR pathway in the TLR4-induced tolerogenic phenotype in human dendritic cells has been reported. PMID: 28256612
TLR4 Asp299Gly polymorphism potentially leading to the development of recurrent hydatidosis, by skewing the immune system towards a Th2 response PMID: 29602972
Study demonstrated that HMGB1 and TLR4 could contribute to the inflammatory lichen planus process in skin. PMID: 29728859
Physical interaction between p38 and eNOS was demonstrated by immunoprecipitation, suggesting a novel, NO-independent mechanism for eNOS regulation of TLR4. In correlation, biopsy samples in patients with systemic lupus erythematous showed reduced eNOS expression with associated elevations in TLR4 and p38, suggesting an in vivo link. PMID: 29061842
The overexpression of miR-140 inhibited the upregulation of the expression of TLR4. PMID: 29901170
TLR4 small interfering RNA blocked hUGT1A1/hNRs downregulation. PMID: 29311138
The expression of TLR4 in perihematoma tissue began to increase within 6 hours after intracerebral hemorrhage and decreased after 72 hours. PMID: 29990607
miR-20a could negatively regulate TLR4 and NLRP3 signaling to protect human aortic endothelial cells from inflammatory injuries. PMID: 29653364
Study provides clear evidence that resistin is a clinically relevant endogenous ligand for TLR4, which promotes tumor progression via TLR4/NF-kappaB/STAT3 signaling. PMID: 28991224
The antitumor effect of curcumin was related to the inhibition HSP70-TLR4 signaling. PMID: 29901164
Activates the NFkappaB pathway through the Tolllike receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/IkappaBalpha axis. PMID: 29916535
An MRP-TLR4 dependent signaling may play an important role in the pathogenesis of autoimmune thyroid diseases. PMID: 29656212
The pathways involved in the effect of ROCK1 in human corneal epithelial cells was preliminarily explained by detecting changes of TLR4-mediated NF-kB and ERK signaling. PMID: 29804125
Serum TLR4 was closely related to AA and associated with some AA-related circulating markers PMID: 29649455
Fibrinogen induced podocyte injury via the TLR4-p38 MAPK-NF-kappaB p65 pathway in focal segmental glomerulosclerosis. PMID: 28407405
Crosstalk between TLR4 and Notch1 signaling regulates the inflammatory response in the IgAN and maybe plays an important role in the progression of IgAN. PMID: 29230705
Data indicate three sites within tenascin-C that directly and cooperatively interact with toll-like receptor 4 (TLR4). PMID: 29150600
Low TLR4 expression is associated with pancreatic ductal adenocarcinoma. PMID: 30246618
An association exists between P. gingivalis and P. intermedia with increased TLR-4 and NF-kappaB expression in the placenta of pre-eclamptic women with periodontitis. PMID: 28349674
TLR4 SNPs were not associated with acute graft rejection in kidney transplant recipients. PMID: 28411360
TLR4 Thr399Ile polymorphism was associated with increased risk of Crohn's disease in Asia and Asians. [meta-analysis] PMID: 29421805
Our results indicated TLR4 SNP rs11536889 may be a marker for intracranial aneurysm risk PMID: 29754966
the expression of TLR4 in gingival tissues and on mast cells increased with the severity of chronic periodontitis, suggesting that TLR4, particularly mast cell TLR4, may be important in the disease process of human chronic periodontitis. PMID: 29488617
Report shows that rheumatoid arthritis (RA) patients express functional TLR4 on peripheral CD8+ T cells that directly promote T-cell function and differentiation to Tc1. The study also suggests that TLR4 signals directly drive Tc1 development and T cell activation independent of TCR engagement. PMID: 28424490
Resistin promoted lung adenocarcinoma metastasis through the TLR4/Src/EGFR/PI3K/NF-kappaB pathway. PMID: 29927028
imcreased TLR4 expression in gastric cardia lesions may be associated with gastric cardia cancer tumorigenesis PMID: 29670922
The immunoenhancement effect of PSP against lung cancer is mediated by TLR4-MAPK/NF-kappaB signaling pathways PMID: 29343453
LPS stimulation induced TLR4 expression and increased pigmentation. TLR4 expression was not detected after single-dose UVA or UVB treatment, but pigmentation increased. Repeated UV treatment induced TLR4 expression and increased pigmentation. LPS stimulation and repeated UV treatment increased IL-6 secretion, and repeated UVB treatment increased IL-10 secretion. PMID: 29063638
data suggest that high-phosphate conditions directly induce vascular calcification via the activation of TLR4/NF-kappaB signaling in VSMCs PMID: 29227975
LncRNA MEG3 ameliorates respiratory syncytial virus infection by suppressing TLR4 signaling. PMID: 29257348
Study shows that the toll-like receptor 4 gene rs1927914 polymorphism was associated with susceptibility to ischemic stroke in males. Moreover, the rs10759932 polymorphism may affect inflammatory response in ischemic stroke patients. PMID: 29075930
There was no difference found in MMP-2, MMP-9 or TLR-4 levels between non-thrombocytopenic and thrombocytopenic septic donors. PLA formation was increased in thrombocytopenic patients. PMID: 29734352
there was a negative correlation between YKL-40 and TLR4 expression in chronic sinusitis patients with nasal polyps. YKL-40 and TLR4 interacted with each other to activate NF-kappaB and promote disease progression. PMID: 29921378
High expression of MMP-9 and TLR4 in patients with COPD may promote inflammatory cell infiltration, induce proliferation of smooth muscle cells, degrade extracellular matrix, and play an important role in lung revascularization. PMID: 28537664
our findings confirm that in south Tunisian patients with IBD, the TLR4-Thr399Ile variant is strongly associated with susceptibility to CD and that the two polymorphisms of this receptor (TLR4-Thr399Ile and TLR4-Asp299Gly) may play a role in the clinical expression of UC. PMID: 29055077
Logistic analysis showed that both rs11536889 and rs7873784 in TLR4 were associated with risk of type-2 diabetes mellitus (T2DM) complicated by tuberculosis (TB) (T2DMTB) in additive and dominant models. Carriers with homozygous and heterozygous mutants of rs11536889 and rs7873784 were associated with higher T2DMTB risk than those with wild-type homozygotes PMID: 29073942
Involvement of the TLR-4 in the mediation of the biglycan action was confirmed using a specific silent agent (siRNA). Taken together, these data could be used to develop new anti-inflammatory approaches PMID: 29339093
TLR4 was significantly up-regulated in synovial tissue samples from rheumatoid arthritis patients. PMID: 28987944
The results revealed a lack of association for TLR4 variant with ischemic stroke and hemorrhagic stroke, although a significant association was observed with the subtypes extracranial large artery. PMID: 28963650
TREM-2 promotes acquired cholesteatoma-induced bone destruction by modulating TLR4 signaling pathway and osteoclasts activation PMID: 27934908

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Subcellular Location

Cell membrane; Single-pass type I membrane protein. Early endosome. Cell projection, ruffle.

Protein Families

Toll-like receptor family

Tissue Specificity

Highly expressed in placenta, spleen and peripheral blood leukocytes. Detected in monocytes, macrophages, dendritic cells and several types of T-cells.

Database Links

HGNC: 11850

OMIM: 603030

KEGG: hsa:7099

STRING: 9606.ENSP00000363089

UniGene: Hs.174312

Code	CSB-E12954h
Size	96T,5×96T,10×96T
Price	Request a Quote or Start an on-line Chat
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Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays): CV%<10%

Three samples of known concentration were tested in twenty assays to assess.