CLIA Kit for Interleukin 28B (IL28B)

IFNL3; Interferon,Lambda 3; Cytokine Zcyto22

Specificity

This assay has high sensitivity and excellent specificity for detection of Interleukin 28B (IL28B).
No significant cross-reactivity or interference between Interleukin 28B (IL28B) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Interleukin 28B (IL28B) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 28B (IL28B) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-104 91
EDTA plasma(n=5) 84-102 98
heparin plasma(n=5) 92-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 28B (IL28B) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Interleukin 28B (IL28B) 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 Interleukin 28B (IL28B) 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) 91-98% 97-104% 78-99% 84-92%
EDTA plasma(n=5) 98-105% 94-105% 96-104% 84-98%
heparin plasma(n=5) 80-95% 78-91% 79-95% 97-105%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Journal of Antimicrobial Chemotherapy Impact of IL28B gene polymorphisms on interferon-λ3 plasma levels during pegylated interferon-α/ribavirin therapy for chronic hepatitis C in patients coinfected with HIV PubMed: 22294646
Cytokine Different effects of three interferons L on Toll-like receptor-related gene expression in HepG2 cells. Pubmed: 24041672
Hepatology Polymorphisms in melanoma differentiation‐associated gene 5 link protein function to clearance of hepatitis C virus Pubmed:25130193
Cytokine. Increased concentration of interferon lambda-3, interferon beta and interleukin-10 in the cerebrospinal fluid of patients with tick-borne encephalitis Pubmed:25461389
J Biomed Res.  Impact of IL28B gene polymorphisms rs8099917 and rs12980275 on response to pegylated interferon-α/ribavirin therapy in chronic hepatitis C genotype 4 … pmc:PMC4726832
Journal of Biomedical Research Impact of IL28B gene polymorphisms rs8099917 and rs12980275 on response to pegylated interferon-α/ribavirin therapy in chronic hepatitis C genotype 4 patients PMC4726832
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