High Sensitive ELISA Kit for Toll Like Receptor 9 (TLR9)

CD289

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive Toll Like Receptor 9 (TLR9).
No significant cross-reactivity or interference between High Sensitive Toll Like Receptor 9 (TLR9) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Toll Like Receptor 9 (TLR9) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Toll Like Receptor 9 (TLR9) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-98 89
EDTA plasma(n=5) 93-101 97
heparin plasma(n=5) 78-93 81

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Toll Like Receptor 9 (TLR9) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Toll Like Receptor 9 (TLR9) 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 High Sensitive Toll Like Receptor 9 (TLR9) 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) 94-105% 82-105% 89-98% 84-101%
EDTA plasma(n=5) 78-101% 82-93% 79-92% 94-102%
heparin plasma(n=5) 85-104% 94-101% 82-96% 78-94%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Chinese Medical Journal Expression of soluble Toll-like receptors in pleural effusions CMJ: 2010823483999506297
DNA Cell Biol. Expression of serum toll-like receptor 9 and oxidative damage markers in benign and malignant breast diseases Pubmed:24906089
Journal of Trace Elements in Medicine and Biology Correlation of serum toll like receptor 9 and trace elements with lipid peroxidation in the patients of breast diseases PubMed: 25744504
Journal of cancer research and clinical oncology TLR9 gene polymorphism− 1486T/C (rs187084) is associated with uterine cervical neoplasm in Mexican female population pubmed:28819773
In Vivo The Relationship Between Sepsis-induced Immunosuppression and Serum Toll-like Receptor 9 Level Pubmed: 30348730
The role of toll-like receptor 9 (TLR9) in Epstein-Barr virus-associated gastric cancer
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