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Human IL-10 QuantiGlo ELISA Kit

Catalog #: Q1000
2 Citations Datasheet / COA / SDS

Discontinued Product

Q1000 has been discontinued.
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human IL-10 QuantiGlo ELISA Kit Summary

Assay Length
5.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (40 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL)
Sensitivity
3.03 pg/mL
Assay Range
3.2 - 10,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human IL-10
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 QuantiGlo Human IL-10 Chemiluminescent Immunoassay is a solid phase ELISA that specifically measures human IL-10 in less than 6 hours. It contains recombinant human IL-10 (expressed in Sf 21 insect cells using a baculovirus expression system) and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-10 showed dose-response curves that are parallel to the standard curves obtained using the recombinant QuantiGlo kit standards. These results indicate that this kit can be used to determine relative mass values for natural IL-10.

Precision

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

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 40 40 40 40
Mean (pg/mL) 25.9 67.1 578 5124 20 48.3 479 3945
Standard Deviation 0.83 1.5 26 204 3.87 4.3 23.3 222
CV% 3.2 2.2 4.5 4 19.4 8.9 4.9 5.6

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 38 38 36 34
Mean (pg/mL) 17.3 57.8 597 5222 17.1 50.3 506 4188
Standard Deviation 0.85 1.85 26.3 67.9 3.35 5.4 32 276
CV% 4.9 3.2 4.4 1.3 19.6 10.7 6.3 6.6

Recovery

The recovery of natural and recombinant human IL-10 spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 101 93-106
EDTA Plasma (n=4) 93 88-99
Heparin Plasma (n=4) 98 92-104
Serum (n=4) 97 90-103

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of IL-10 were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Scientific Data

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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 -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: IL-10

Interleukin-10 functions as an anti-inflammatory cytokine by inhibiting the expansion and activation of Th1 cells and Th17 cells and by promoting the development of M2 macrophages and regulatory T cells (Treg). Within a tumor microrenvironment, however, IL-10 can inhibit the expansion of both Treg and myeloid-derived suppressor cells (MDSC). IL-10 exerts protective effects including limiting tissue damage in arthritic inflammation and promoting muscle regeneration after injury, but it also contributes to the persistence of viral infections. IL-10 signals through a receptor complex composed of IL-10 R alpha and IL-10 R beta. IL-10 R beta additionally associates with IL-20 R alpha, IL-22 R alpha 1, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.

Long Name:
Interleukin 10
Entrez Gene IDs:
3586 (Human); 16153 (Mouse); 25325 (Rat); 397106 (Porcine); 403628 (Canine); 102133450 (Cynomolgus Monkey); 493683 (Feline); 100715618 (Guinea Pig); 2949786 (Viral)
Alternate Names:
CSIF; CSIFMGC126450; Cytokine synthesis inhibitory factor; GVHDS; IL10; IL-10; IL10A; IL-10MGC126451; interleukin 10; interleukin-10; TGIF

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. 50 µL Assay Diluent
  3.   Add 50 µL of Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20-40 minutes on the benchtop.
  9.   Determine the RLU of each well using a luminometer.

Citations for Human IL-10 QuantiGlo 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.

2 Citations: Showing 1 - 2
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  1. Human interleukin-10 gene transfer is protective in a rat model of Parkinson's disease.
    Authors: Johnston LC, Su X, Maguire-Zeiss K, Horovitz K, Ankoudinova I, Guschin D, Hadaczek P, Federoff HJ, Bankiewicz K, Forsayeth J
    Mol. Ther., 2008-06-10;16(8):1392-9.
    Species: Human
    Sample Types: Tissue Homogenates
  2. Redirection of human autoreactive T-cells Upon interaction with dendritic cells modulated by TX527, an analog of 1,25 dihydroxyvitamin D(3).
    Authors: van Halteren AG, van Etten E, de Jong EC, Bouillon R, Roep BO, Mathieu C
    Diabetes, 2002-07-01;51(7):2119-25.
    Species: Human
    Sample Types: Cell Culture Supernates

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