ELISA Kit for Trefoil Factor 3 (TFF3)

HITF; ITF3; TFI; HP1.B; Trefoil Factor 3, Intestinal

Specificity

This assay has high sensitivity and excellent specificity for detection of Trefoil Factor 3 (TFF3).
No significant cross-reactivity or interference between Trefoil Factor 3 (TFF3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Trefoil Factor 3 (TFF3) and the recovery rates were calculated by comparing the measured value to the expected amount of Trefoil Factor 3 (TFF3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-103 92
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 78-89 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Trefoil Factor 3 (TFF3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Trefoil Factor 3 (TFF3) 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 Trefoil Factor 3 (TFF3) 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) 84-96% 87-95% 92-104% 85-95%
EDTA plasma(n=5) 98-105% 96-104% 80-99% 89-96%
heparin plasma(n=5) 78-91% 85-103% 96-104% 96-105%

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
BMC Gastroenterology Serum trefoil factor 3 is a promising non-invasive biomarker for gastric cancer screening: A monocentric cohort study in China Biomedcentral: Source
Technology in Cancer Research & Treatment Serum Trefoil Factor 3 as a Protein Biomarker for the Diagnosis of Colorectal Cancer. pubmed:27760866
PLoS One Obstructive sleep apnea and rhonchopathy are associated with downregulation of trefoil factor family peptide 3 (TFF3)-Implications of changes in oral mucus composition. pubmed:29028798
Microbial Pathogenesis Protective effects of sinomenine against LPS-induced inflammation in piglets. pubmed:28757275
Scientific Reports Nucleotide-mediated SPDEF modulates TFF3-mediated wound healing and intestinal barrier function during the weaning process Pubmed:29555969
Scientific Reports Kidney-based in vivo model for drug-induced nephrotoxicity testing Pubmed: 32796873
International Journal of Molecular Sciences Human Synovia Contains Trefoil Factor Family (TFF) Peptides 1–3 Although Synovial Membrane Only Produces TFF3: Implications in Osteoarthritis and Rheumatoid …
Int. J. Mol. Sci. Untersuchungen der Trefoil factor family Peptide 1-3 in humaner Synovialfl¨¹ssigkeit und Synovialmembran und deren Bedeutung bei Arthrose und?¡­
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