Human TFF3 Quantikine ELISA Kit

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DTFF30
Control Products Available
Human TFF3 ELISA Standard Curve
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Procedure
Citations (5)
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Human TFF3 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (10 uL)
Sensitivity
13.1 pg/mL
Assay Range
39.0 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human Trefoil Factor 3 (TFF3)
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human TFF3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human TFF3 in cell culture supernates, serum, plasma, saliva, and urine. It contains CHO cell-expressed recombinant human TFF3 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TFF3 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human TFF3.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of a known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in separate assays to determine inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 4 4 4 4 4 4
Mean 276 849 1684 286 894 1712
Standard Deviation 6.01 9.13 37.4 18.3 44 95.9
CV% 2.2 1.1 2.2 6.4 4.9 5.6

Recovery

The recovery of human TFF3 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 100 90-114

Linearity

To assess the linearity of the assay, samples containing high concentrations of human TFF3 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human TFF3 ELISA Linearity

Data Examples

Human TFF3 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TFF3

TFF3 (Trefoil Factor 3), also known as ITF and P1.B, is a peptide that plays an important role in epithelial regeneration and wound healing. It can form disulfide-linked dimers or associate into disulfide-linked complexes with the intestinal mucous proteins FCGBP and MUC-2. TFF3 is expressed by epithelial goblet cells in the respiratory tract, biliary and breast ducts, small and large intestine, and cardia of the stomach as well as by chondrocytes during bone development. TFF3 is upregulated in response to gastrointestinal epithelial disruptions and promotes epithelial wound healing. It enhances tumor cell invasion and metastasis by supporting hypoxia-induced VEGF upregulation.

Long Name:
Trefoil Factor 3
Entrez Gene IDs:
7033 (Human); 21786 (Mouse); 25563 (Rat)
Alternate Names:
HITF, human intestinal trefoil factor10Intestinal trefoil factor; ITF; TFF3; TFI; trefoil factor 3 (intestinal)
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human TFF3 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Serum trefoil factor 3 predicts disease activity in patients with ulcerative colitis
    Authors: R Nakov, T Velikova, V Nakov, G Ianiro, V Gerova, L Tankova
    Eur Rev Med Pharmacol Sci, 2019;23(2):788-794.
    Species: Human
    Sample Types: Serum
  2. Urinary Cysteine-Rich Protein 61 and Trefoil Factor 3 as Diagnostic Biomarkers for Colorectal Cancer
    Authors: T Shimura, H Iwasaki, M Kitagawa, M Ebi, T Yamada, T Yamada, T Katano, H Nisie, Y Okamoto, K Ozeki, T Mizoshita, H Kataoka
    Transl Oncol, 2019;12(3):539-544.
    Species: Human
    Sample Types: Urine
  3. Trefoil Factor 3 (TFF3) Is Involved in Cell Migration for Skeletal Repair
    Authors: K Krüger, S Schmid, F Paulsen, A Ignatius, P Klinger, T Hotfiel, B Swoboda, K Gelse
    Int J Mol Sci, 2019;20(17):.
    Species: Human
    Sample Types: Serum
  4. Pharmacological Inhibition of TFF3 Enhances Sensitivity of CMS4 Colorectal Carcinoma to 5-Fluorouracil through Inhibition of p44/42 MAPK
    Authors: RM Chen, YS Chiou, QY Chong, HM Poh, TZ Tan, MY Zhang, L Ma, T Zhu, V Pandey, Basappa, AP Kumar, PE Lobie
    Int J Mol Sci, 2019;20(24):.
    Species: Xenograft
    Sample Types: Serum
  5. Trefoil Factor 1 Excretion Is Increased in Early Stages of Chronic Kidney Disease.
    Authors: Lebherz-Eichinger D, Tudor B, Ankersmit H, Reiter T, Haas M, Roth-Walter F, Krenn C, Roth G
    PLoS ONE, 2015;10(9):e0138312.

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