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hIL-10 QKit (1 Kit)
品牌： R&D Systems
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Product Summary
The Quantikine Human IL-10 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-10 in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human IL-10 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-10 and does not cross-react with viral IL-10. Results obtained using natural human IL-10 showed linear curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate this kit can be used to determine relative mass values for natural human IL-10.
Sample Values
Serum/Plasma - Forty serum and plasma samples from apparently healthy volunteers were evaluated for the presence of human IL-10 in this assay. All samples measured less than the lowest standard, 7.8 pg/mL. No medical histories were available for the donors used in this study.Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 units/mL penicillin, 100 μg/mL streptomycin sulfate. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1, 3, and 5 days. Aliquots of the cell culture supernates were removed and assayed for human IL-10.
| Condition | Day 1 (pg/mL) | Day 3 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 98 | 134 | 127 |
| Stimulated | 2756 | 3563 | 2255 |
Recovery
The recovery of human IL-10 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 98 | 88-110 |
| Citrate Plasma (n=5) | 96 | 87-108 |
| EDTA Plasma (n=5) | 92 | 75-105 |
| Heparin Plasma (n=5) | 94 | 84-105 |
| Serum (n=5) | 100 | 92-110 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-10 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, working standards, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.
- Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well, and cover with a new plate sealer.
- For Serum & Plasma Samples: Incubate at room temperature for 2 hours.
For Cell Culture Supernate Samples: Incubate at room temperature for 1 hour. - Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. PROTECT FROM LIGHT.
- For Serum & Plasma Samples: Incubate at room temperature for 30 minutes.
For Cell Culture Supernate Samples: Incubate at room temperature for 20 minutes. - 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.
50 µL Assay Diluent for Serum & Plasma Samples Only
200 µL Standard, control, or sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Human IL-10 Quantikine ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Citrate Plasma (200 uL)
Sensitivity
3.9 pg/mL
Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate 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.
背景
别名
CSIF,CSIFMGC126450,Cytokine synthesis inhibitory factor,GVHDS,IL10,IL-10,IL10A,IL-10MGC126451,interleukin 10,interleukin-10,TGIF,Interleukin 10
背景
Background: IL-10
Interleukin-10 (IL-10), also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL-10 alpha -helical cytokine family that also includes IL-19, IL-20, IL-22, IL-24, and IL-26/AK155 (1-3). IL-10 is secreted by many activated hematopoietic cell types as well as hepatic stellate cells, keratinocytes, and placental cytotrophoblasts. Whereas human IL-10 is active on mouse cells, mouse IL-10 does not act on human cells (4, 5). Mature human IL-10 shares 86% amino acid sequence identity with equine IL-10 and 72% - 80% with bovine, canine, feline, guinea pig, mouse, ovine, porcine, and rat IL-10. It contains two intrachain disulfide bridges and is expressed as a 36 kDa noncovalently-associated homodimer (4, 6, 7).
IL-10 mediates its biological activities through a heteromeric receptor complex composed of the type II cytokine receptor subunits IL-10 R alpha and IL-10 R beta. IL-10 R alpha is a 110 kDa transmembrane glycoprotein that is expressed on lymphocytes, NK cells, macrophages, monocytes, astrocytes, intestinal epithelial cells, cytotrophoblasts, and activated hepatic stellate cells (8-13), while the 75 kDa transmembrane IL-10 R beta is widely expressed (14, 15). The IL-10 dimer binds to two IL-10 R alpha chains, triggering recruitment of two IL-10 R beta chains (14, 15). IL-10 R beta does not bind IL-10 directly but is required for signal transduction. IL-10 R beta also 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 (16-18).
The involvement of IL-10 in immunoregulation includes both suppressive and stimulatory effects. It functions as an anti-inflammatory cytokine by inhibiting the expansion and activation of Th1 cells and Th17 cells (19-21) and by promoting the development of M2 macrophages (21). Its expression by immunosuppressive regulatory T cells (Treg) and regulatory B cells is important for Treg proliferation (19). Within a tumor microrenvironment, however, IL-10 inhibits the expansion of Treg as well as myeloid-derived suppressor cells (22, 23). IL-10 induces the intratumoral accumulation and activation of CD8+ T cells (24, 25). IL-10 exerts protective effects including limiting tissue damage in arthritic inflammation (19) and promoting muscle regeneration after injury (21), but it also contributes to the persistence of viral infections (26). The levels of IL-10 are elevated in Sjogren's syndrome (saliva), primary CNS lymphoma (cerebrospinal fluid), and ovarian cancer (serum and ascites) (27-29). Its levels are decreased in the serum in patients with recurrent heart attacks or during preeclampsia and also in the seminal fluid of infertile men (30-32).
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
研究领域
制备和贮存
保存方式
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.
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