ELISA Kit for Interleukin 32 (IL32)

NK4; TAIF; TAIFb; TAIFd; Natural Killer Cell Transcript 4; Natural killer cells protein 4; Tumor necrosis factor alpha-inducing factor

Specificity

This assay has high sensitivity and excellent specificity for detection of Interleukin 32 (IL32).
No significant cross-reactivity or interference between Interleukin 32 (IL32) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Interleukin 32 (IL32) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 32 (IL32) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-103 94
EDTA plasma(n=5) 90-104 96
heparin plasma(n=5) 95-103 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 32 (IL32) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Interleukin 32 (IL32) 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 32 (IL32) 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) 90-101% 85-102% 78-95% 81-94%
EDTA plasma(n=5) 87-95% 78-99% 95-104% 95-102%
heparin plasma(n=5) 93-101% 78-97% 82-96% 86-105%

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
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
Lupus. Three cases of lupus nephritis patients with serum interleukin-32γ detection Pubmed:24879659
Tumour Biol Clinical significance of serum interleukin-29, interleukin-32, and tumor necrosis factor alpha levels in patients with gastric cancer PubMed: 26219901
Biomedical Reports Calprotectin in serum and zonulin in serum and feces are elevated after introduction of a diet with lower carbohydrate content and higher fiber, fat and protein contents. pubmed:28413639
Arab Journal of Gastroenterology Role of several cytokines and adhesion molecules in the diagnosis and prediction of survival ofhepatocellular carcinoma. pubmed:27916547
ResearchGate SERUM INTERLEUKIN-32 (IL-32) LEVELS MAY HAVE DIAGNOSTIC AND PROGNOSTIC ROLES IN PATIENTS WITH... doi:10.19193/0393-6384_2017_4_091
Biomedical Reports Calprotectin in serum and zonulin in serum and feces are elevated after introduction of a diet with lower carbohydrate content and higher fiber, fat and protein contents 10.3892/br.2017.865
Journal of Diabetes & Metabolism High Fiber Fat and Protein Contents Lead to Increased Satiety Reduced Sweet Cravings and Decreased Gastrointestinal Symptoms Independently of Anthropometric Hormonal and Metabolic Factors 10.4172/2155-6156.1000733 
IL-31, IL-32 and IL-33 may Serve as Diagnosis Biomarkers in Gastric Cancer
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