Human TSLP Quantikine ELISA Kit

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DTSLP0
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Human TSLP ELISA Standard Curve
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Human TSLP 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 (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
9.87 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human TSLP
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 TSLP immunoassay is a 4.5 hour sandwich-type solid phase ELISA designed to measure human TSLP in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human TSLP and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TSLP 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 TSLP.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 193 729 1318 230 775 1341
Standard Deviation 17.9 52.8 108 21.9 53.4 80.6
CV% 9.2 7.2 8.2 9.5 6.9 6

Recovery

The recovery of TSLP spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 90-107
EDTA Plasma (n=4) 96 81-105
Heparin Plasma (n=4) 97 91-104
Serum (n=4) 94 88-101

Linearity

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

Data Examples

Human TSLP 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: TSLP

Thymic Stromal Lymphopoietin (TSLP) was originally identified as an activity from the conditioned medium of a mouse thymic stromal cell line that promoted the development of B cells. The activities of mouse TSLP overlap with, but are distinct from, those of mouse IL-7. Both mouse TSLP and IL-7 can co-stimulate growth of thymocytes and mature T cells, and support B lymphopoiesis in long-term cultures of fetal liver cells and bone-marrow cells. Whereas mouse IL-7 facilitates the development of B220+/IgM- pre-B cells, mouse TSLP promotes the development B220+/IgM+ B cells. Human TSLP was reported to preferentially stimulate myeloid cells, inducing the release of T cell-attracting chemokines from monocytes and enhancing the maturation of CD11c+ dendritic cells. Human TSLP cDNA encodes a 159 amino acid (aa) residue precursor protein with a 28 aa signal sequence. Within the mature region, six of the seven cysteine residues present in the mouse TSLP involved in intramolecular disulfide bond formation are conserved in the human TSLP. Human TSLP shares approximately 43% aa sequence identity with mouse TSLP. By Northern blot analysis, human TSLP expression has been detected in many tissues with the highest expressions in heart, liver, testis and prostate. TSLP signals through a heterodimeric receptor complex that consists of IL-7 R alpha and the TSLP R, a member of the hemopoietin receptor family most closely related to Rgammac.

TSLP R, also named Delta and CRLM-2 (Cytokine Receptor-like Module-2), was originally cloned as a novel type 1 cytokine receptor with similarity to the common gamma chain (gamma c). It was subsequently identified as a subunit of the cellular receptor for the IL-7-like cytokine TSLP and termed TSLP R. TSLP R shows 24% sequence identity with the mouse gc receptor. TSLP R expression is constitutive in immune and hematopoietic cells, but is up-regulated in Th2 cells.

Long Name:
Thymic Stromal Lymphopoietin
Entrez Gene IDs:
85480 (Human); 53603 (Mouse); 102119254 (Cynomolgus Monkey)
Alternate Names:
thymic stromal lymphopoietin; TSLP
⚠ 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.
  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.
  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. 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 TSLP 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.

13 Citations: Showing 1 - 10
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  1. IL-17A-associated IKK-? signaling induced TSLP production in epithelial cells of COPD patients
    Authors: G Anzalone, GD Albano, AM Montalbano, L Riccobono, A Bonanno, R Gagliardo, F Bucchieri, R Marchese, M Moscato, M Profita
    Exp. Mol. Med., 2018;50(10):131.
    Species: Human
    Sample Types: Sputum Supernates
  2. Estrogen restricts the apoptosis of endometrial stromal cells by promoting TSLP secretion
    Authors: HL Yang, KK Chang, J Mei, WJ Zhou, LB Liu, L Yao, Y Meng, MY Wang, SY Ha, ZZ Lai, JF Ye, DJ Li, MQ Li
    Mol Med Rep, 2018;0(0):.
  3. Cytokine Profile of Patients with Allergic Rhinitis Caused by Pollen, Mite, and Microbial Allergen Sensitization
    Authors: YA Tyurin, SA Lissovskay, RS Fassahov, IG Mustafin, AF Shamsutdin, MA Shilova, AA Rizvanov
    J Immunol Res, 2017;2017(0):3054217.
    Species: Human
    Sample Types: Serum
  4. Betamethasone Butyrate Propionate Inhibits the Induction of Thymic Stromal Lymphopoietin in Cultured Normal Human Keratinocytes
    Authors: Wei Zhang
    Ann Dermatol, 2016;28(6):772-775.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. TNF-alpha and Th2 cytokines induce atopic dermatitis-like features on epidermal differentiation proteins and stratum corneum lipids in human skin equivalents.
    Authors: Danso M, van Drongelen V, Mulder A, van Esch J, Scott H, van Smeden J, El Ghalbzouri A, Bouwstra J
    J Invest Dermatol, 2014;134(7):1941-50.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. MicroRNA-375 regulation of thymic stromal lymphopoietin by diesel exhaust particles and ambient particulate matter in human bronchial epithelial cells.
    Authors: Bleck B, Grunig G, Chiu A, Liu M, Gordon T, Kazeros A, Reibman J
    J Immunol, 2013;190(7):3757-63.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Infliximab infusions for netherton syndrome: sustained clinical improvement correlates with a reduction of thymic stromal lymphopoietin levels in the skin.
    Authors: Fontao L, Laffitte E, Briot A, Kaya G, Roux-Lombard P, Fraitag S, Hovnanian AA, Saurat JH
    J. Invest. Dermatol., 2011;131(9):1947-50.
    Species: Human
    Sample Types: Serum
  8. Opposing roles of IL-17A and IL-25 in the regulation of TSLP production in human nasal epithelial cells.
    Authors: Xu G, Zhang L, Wang DY, Xu R, Liu Z, Han DM, Wang XD, Zuo KJ, Li HB
    Allergy, 2010;65(5):581-9.
    Species: Human
    Sample Types: Nasal Lavage Fluid
  9. TSLP and downstream molecules in experimental mouse allergic conjunctivitis.
    Authors: Zheng X, Ma P, De Paiva CS, Cunningham MA, Hwang CS, Pflugfelder SC, Li DQ
    Invest. Ophthalmol. Vis. Sci., 2010;51(6):3076-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Thymic stromal lymphopoietin-induced human asthmatic airway epithelial cell proliferation through an IL-13-dependent pathway.
    Authors: Semlali A, Jacques E, Koussih L, Gounni AS, Chakir J
    J. Allergy Clin. Immunol., 2010;125(4):844-50.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Human corneal epithelium-derived thymic stromal lymphopoietin links the innate and adaptive immune responses via TLRs and Th2 cytokines.
    Authors: Ma P, Bian F, Wang Z, Zheng X, Chotikavanich S, Pflugfelder SC, Li DQ
    Invest. Ophthalmol. Vis. Sci., 2009;50(6):2702-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Inhibition of double-stranded RNA-induced TSLP in human keratinocytes by glucocorticoids.
    Authors: Le TA, Takai T, Kinoshita H, Suto H, Ikeda S, Okumura K, Ogawa H
    Allergy, 2009;64(8):1231-2.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Allergen inhalation decreases adenosine receptor expression in sputum and blood of asthma patients.
    Authors: Versluis M, van den Berge M, Timens W, Luijk B, Rutgers B, Lammers JW, Postma DS, Hylkema MN
    Allergy, 2008;63(9):1186-94.
    Species: Human
    Sample Types: Sputum

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