Multiplex Assay Kit for Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay)

CD282; TIL4; Toll/interleukin-1 receptor-like protein 4

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-103 95
EDTA plasma(n=5) 83-97 88
heparin plasma(n=5) 98-105 102
sodium citrate plasma(n=5) 88-105 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Toll Like Receptor 2 (TLR2) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 82-103% 79-94% 84-97% 93-102%
EDTA plasma(n=5) 85-104% 86-94% 88-97% 98-105%
heparin plasma(n=5) 97-105% 97-104% 83-101% 85-94%
sodium citrate plasma(n=5) 78-93% 97-105% 78-105% 85-102%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:TLR2) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Chinese Medical Journal Expression of soluble Toll-like receptors in pleural effusions CMJ: 2010823483999506297
PLoS One Baicalein Reduces Airway Injury in Allergen and IL-13 Induced Airway Inflammation PubMed: PMC3639905
Clinical and Developmental Immunology TLR2 in Pleural Fluid Is Modulated by Talc Particles during Pleurodesis PubMed: PMC3529880
Advances in Virology The Possible Role of TLR2 in Chronic Hepatitis B Patients with Precore Mutation Pubmed: 24187552
J Matern Fetal Neonatal Med. Cervical and vaginal fluid soluble Toll-like receptor 2 in pregnancies complicated by preterm prelabor rupture of membranes Pubmed:25028801
Respiratory Virology and Immunogenicity Advances in Experimental Medicine and Biology Cytokines and Toll-Like Receptors in the Immune Response to Influenza Vaccination Pubmed:25248345
Iran J Immunol. Correlation?between?salivary?Toll like receptor-2?concentration?and early childhood caries. Pubmed:25265998
Int J Dermatol. Membrane and soluble Toll‐like receptor 2 in patients with psoriasis treated by Goeckerman therapy Pubmed:25266302
BMC Immunology The discriminative capacity of soluble Toll-like receptor (sTLR)2 and sTLR4 in inflammatory diseases Pubmed:25406630
Pathol Biol (Paris) Cathelicidin related antimicrobial peptide, laminin, Toll-like receptors and chemokines levels in experimental hypersensitivity pneumonitis in mice PubMed: 25834936
Plos One Soluble Toll-Like Receptors 2 and 4 in Cerebrospinal Fluid of Patients with Acute Hydrocephalus followingAneurysmal Subarachnoid Haemorrhage. Pubmed:27223696
Open Forum Infect Dis The Effect of Rosuvastatin on Markers of Immune Activation in Treatment-Naive Human Immunodeficiency Virus-Patients Pubmed:26835476
Perfusion Pentraxin 3 and other inflammatory biomarkers related to atrial fibrillation in cardiac surgery pubmed:27852958
Journal For Neurovirology Soluble TLR2 and 4 concentrations in cerebrospinal fluid in HIV/SIV-relatedneuropathological conditions. pubmed:27882497
Acta Orthopaedica Toll-like receptors and their soluble forms differ in the knee and thumb basal osteoarthritic joints pubmed:28093922
Biological Markers and Guided Therapy Features of the Pattern Recognition Receptor TLR-2 Expression and Cytokines IL-2, IL-4 Profile in Children with Food Hypersensitivity 10.12988:bmgt.2018.812
CHILD`S HEALTH Prospective study on the significance of mucosal barrier dysfunction markers in assessing the effectiveness of the new strategy for treating food hypersensitivity in … 10.22141:2224-0551.13.2.2018.129548
Neurological Sciences Vinpocetine regulates levels of circulating TLRs in Parkinson's disease patients Pubmed: 30315378
Biological Markers in Fundamental and Clinical Medicine PROGNOSTIC ROLE OF IL-4, IL-2 AND TLR-2 FOR PREDICTING OF COURSE OF FOOD ALLERGY IN CHILDREN Doi: 10.29256/v.02.02.2018.escbm71
Biological Markers in Fundamental and Clinical Medicine STUDY OF THE ACTION OF CHROMIUM-CONTAINING BIOPREPARATIONS FROM SPIRULINA ON INSULIN ACTIVITY IN RATS Doi: 10.29256/v.02.02.2018.escbm72
Biomedicine & Pharmacotherapy Opuntioside, opuntiol and its metallic nanoparticles attenuate adjuvant-induced arthritis: Novel suppressors of Toll-like receptors-2 and-4
PLoS One Aberrant DNA methylation of the toll-like receptors 2 and 6 genes in patients with obstructive sleep apnea Pubmed: 32069296
Georgian medical news ASSOCIATIONS BETWEEN EFFICACY OF THE THERAPY AND CIRCADIAN FLUCTUATIONS OF ENDOTHELIAL NITRIC OXIDE SYNTHASE WITH TOLL-LIKE … Pubmed: 32242853
Medical and Clinical Chemistry ДОбОвА вАРІАбЕЛЬНІСТЬ РІвНЯ РОзЧИННИХ ТОЛ-пОДІбНИХ РЕцЕпТОРІв 2 в СИРОвАТцІ КРОвІ ЖІНОК, ХвОРИХ НА РЕвМАТОїДНИй АРТРИТ: зв' …
Sciences of Europe FEATURES OF MYOCARDIAL REMODELING IN MEN WITH POST-INFARCTION CARDIOSCLEROSIS AND ITS RELATIONSHIP WITH TOLL-LIKE …
CLINICAL INVESTIGATIONS Levels of toll-like receptors 2 and 4 in serum of men with postinfarction cardiosclerosis and its relationship with antiphospholipid syndrome components
Acta Obstet Gynecol Scand A value of soluble Toll‐like receptor 2 and 4 in vaginally obtained amniotic fluid for the prediction of histological chorioamnionitis 34244993
Clinical and Translational Allergy Altered innate immune profile in blood of systemic mastocytosis patients Pubmed:35734269
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