Human TSLP Quantikine ELISA Kit

  (11 citations)     
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Assay Procedure
Citations (11)
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  • 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.
Control Available
QC178 , Quantikine Immunoassay Control Set 571 for Human TSLP - Please Inquire
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.
 TSLP [HRP]
Product Datasheets

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Preparation and Storage
  • 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)
  • Alternate Names:
    thymic stromal lymphopoietin; TSLP
Related Research Areas
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:

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.

11 Citations: Showing 1 - 10
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Species
Sample Type
  1. 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 Type: Serum
  2. 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 Type: Cell Culture Supernates
  3. 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 Type: Cell Culture Supernates
  4. 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 Type: Cell Culture Supernates
  5. 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 Type: Serum
  6. 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 Type: Nasal Lavage
  7. 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 Type: Cell Culture Supernates
  8. 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 Type: Cell Culture Supernates
  9. 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 Type: Cell Culture Supernates
  10. 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 Type: Cell Culture Supernates
  11. 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 Type: Sputum
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Quantikine Immunoassay Control Set 571 for Human TSLP

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Quantikine ELISA Wash Buffer 1

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