Rat IL-10 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY522
DY522-05
Ancillary Products Available
Rat IL-10 ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (29)
FAQs
Supplemental Products
Reviews (1)

Rat IL-10 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat IL-10. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

 

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

 

Data Example

Rat IL-10 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.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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 rat IL-10 shares 85% amino acid sequence identity with mouse IL-10 and 71% - 79% with bovine, canine, equine, feline, guinea pig, human, ovine, and porcine 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 microenvironment, 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 have been found to be elevated in Sjogren's syndrome (saliva), primary CNS lymphoma (cerebrospinal fluid), and ovarian cancer (serum and ascites) (27-29). Its levels have been found to be decreased in the serum of 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); 493683 (Feline); 100135553 (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

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Rat IL-10 DuoSet ELISA

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.

29 Citations: Showing 1 - 10
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  1. Over-Activated Proteasome Mediates Neuroinflammation on Acute Intracerebral Hemorrhage in Rats
    Authors: HK Liew, WF Hu, PB Lin, PK Wang, AP Tsai, CY Pang, TY Chen
    Cells, 2019;8(11):.
    Species: Rat
    Sample Types: Tissue Lysates
  2. Protective effect of Platymiscium floribundum Vog. in tree extract on periodontitis inflammation in rats
    Authors: JMO Freire, HV Chaves, AH Teixeira, LHT de Sousa, IR Pinto, JJDN Costa, NA de Sousa, KMA Pereira, VCC Girão, VCS Ferreira, JEÁ Dos Santos, MAS Lima, ATA Pimenta, RC Montenegro, MEA de Moraes, VPT Pinto, GC Filho, MM Bezerra
    PLoS ONE, 2019;14(11):e0223800.
    Species: Rat
    Sample Types: Gingival Fluid
  3. The Preventive Effects of Greenshell Mussel (Perna canaliculus) on Early-Stage Metabolic Osteoarthritis in Rats with Diet-Induced Obesity
    Authors: P Siriarchav, MC Kruger, MR Miller, HS Tian, FM Wolber
    Nutrients, 2019;11(7):.
    Species: Rat
    Sample Types: Plasma
  4. Polyphenols-Rich Fruit (Euterpe edulis Mart.) Prevents Peripheral Inflammatory Pathway Activation by the Short-Term High-Fat Diet
    Authors: AB Santamarin, G Jamar, LV Mennitti, DA Ribeiro, CM Cardoso, VV de Rosso, LM Oyama, LP Pisani
    Molecules, 2019;24(9):.
    Species: Rat
    Sample Types: Tissue Homogenates
  5. Hyperbilirubinemia in Gunn Rats is Associated with Decreased Inflammatory Response in LPS-Mediated Systemic Inflammation
    Authors: P Valaskova, A Dvorak, M Lenicek, K Zizalova, N Kutinova-C, J Zelenka, M Cahova, L Vitek, L Muchova
    Int J Mol Sci, 2019;20(9):.
    Species: Rat
    Sample Types: Serum
  6. Cholinergic Stimulation by Pyridostigmine Bromide Before Myocardial Infarction Prevent Cardiac and Autonomic Dysfunction
    Authors: CA Barboza, AR Fukushima, N Carrozzi, JF Machi, PMM Dourado, CT Mostarda, MC Irigoyen, L Nathanson, M Morris, EC Caperuto, B Rodrigues
    Sci Rep, 2019;9(1):2481.
    Species: Rat
    Sample Types: Plasma
  7. Oral neonatal antibiotic treatment perturbs gut microbiota and aggravates central nervous system autoimmunity in Dark Agouti rats
    Authors: S Stanisavlj, A ?epi?, S Boji?, K Veljovi?, S Mihajlovi?, N ?edovi?, B Jevti?, M Mom?ilovi?, M Lazarevi?, M Mostarica, ? Miljkovi?, N Goli?
    Sci Rep, 2019;9(1):918.
    Species: Rat
    Sample Types: Cell Culture Supernates
  8. Leukemia inhibitory factor modulates the peripheral immune response in a rat model of emergent large vessel occlusion
    Authors: SM Davis, LA Collier, ED Winford, CC Leonardo, CT Ajmo, EA Foran, TJ Kopper, JC Gensel, KR Pennypacke
    J Neuroinflammation, 2018;15(1):288.
    Species: Rat
    Sample Types: Tissue Homogenates
  9. Lactobacillus fermentum HP3-Mediated Fermented Hericium erinaceus Juice as a Health Promoting Food Supplement to Manage Diabetes Mellitus
    Authors: C Chaiyasut, S Woraharn, BS Sivamaruth, N Lailerd, P Kesika, S Peerajan
    J Evid Based Integr Med, 2018;23(0):2515690X18765.
    Species: Rat
    Sample Types: Serum
  10. Early exposure to distinct sources of lipids affects differently the development and hepatic inflammatory profiles of 21-day-old rat offspring
    Authors: LV Mennitti, LM Oyama, AB Santamarin, CMDPO do Nascime, LP Pisani
    J Inflamm Res, 2018;11(0):11-24.
    Species: Rat
    Sample Types: Tissue Homogenates
  11. Characterization of the Myocardial Inflammatory Response in Acute Stress-Induced (Takotsubo) Cardiomyopathy
    Authors: HM Wilson, L Cheyne, PAJ Brown, K Kerr, A Hannah, J Srinivasan, N Duniak, G Horgan, DK Dawson
    JACC Basic Transl Sci, 2018;3(6):766-778.
    Species: Rat
    Sample Types: Serum
  12. Obesity and Type 2 Diabetes mellitus induce lipopolysaccharide tolerance in rat neutrophils
    Authors: WMT Kuwabara, CNF Yokota, R Curi, TC Alba-Loure
    Sci Rep, 2018;8(1):17534.
    Species: Rat
    Sample Types: BALF
  13. The Use of Ju�ara (Euterpe edulis Mart.) Supplementation for Suppression of NF-?B Pathway in the Hypothalamus after High-Fat Diet in Wistar Rats
    Authors: AB Santamarin, G Jamar, LV Mennitti, VV de Rosso, HC Cesar, LM Oyama, LP Pisani
    Molecules, 2018;23(7):.
    Species: Rat
    Sample Types: Tissue Homogenates
  14. Human Mesenchymal Stem Cell Secretome from Bone Marrow or Adipose-Derived Tissue Sources for Treatment of Hypoxia-Induced Pulmonary Epithelial Injury
    Authors: N Shologu, M Scully, JG Laffey, D O'Toole
    Int J Mol Sci, 2018;19(10):.
    Species: Rat
    Sample Types: Cell Culture Supernates
  15. Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTP? receptors promotes a beneficial inflammatory response following spinal cord injury
    Authors: S Dyck, H Kataria, A Alizadeh, KT Santhosh, B Lang, J Silver, S Karimi-Abd
    J Neuroinflammation, 2018;15(1):90.
    Species: Rat
    Sample Types: Cell Culture Supernates
  16. The Major Histocompatibility Complex Class III Haplotype Ltab-Ncr3 Regulates Adjuvant-Induced but Not Antigen-Induced Autoimmunity
    Authors: AC Yau, J Tuncel, R Holmdahl
    Am. J. Pathol, 2017;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  17. Incline treadmill exercise suppresses pain hypersensitivity associated with the modulation of pro-inflammatory cytokines and anti-inflammatory cytokine in rats with peripheral nerve injury
    Authors: KL Tsai, PC Huang, LK Wang, CH Hung, YW Chen
    Neurosci. Lett, 2017;643(0):27-31.
    Species: Rat
    Sample Types: Tissue Homogenates
  18. Different Dietary Proportions of Fish Oil Regulate Inflammatory Factors but Do Not Change Intestinal Tight Junction ZO-1 Expression in Ethanol-Fed Rats
    Authors: YW Chien, HC Peng, YL Chen, MH Pai, HY Wang, HL Chuang, SC Yang
    Mediators Inflamm., 2017;2017(0):5801768.
    Species: Rat
    Sample Types: Tissue Homogenates
  19. Impact of gestational nicotine exposure on intrauterine and fetal infection in a rodent model
    Authors: M von Chamie, L Reyes, LF Hayward, MB Brown
    Biol. Reprod., 2017;96(5):1071-1084.
    Species: Rat
    Sample Types: Whole Tissue
    Applications: ELISA Capture
  20. Food-grade TiO2 impairs intestinal and systemic immune homeostasis, initiates preneoplastic lesions and promotes aberrant crypt development in the rat colon
    Authors: S Bettini, E Boutet-Rob, C Cartier, C Com‚ra, E Gaultier, J Dupuy, N Naud, S Tach‚, P Grysan, S Reguer, N Thieriet, M R‚fr‚giers, D ThiaudiŠre, JP Cravedi, M CarriŠre, JN Audinot, FH Pierre, L Guzylack-P, E Houdeau
    Sci Rep, 2017;7(0):40373.
    Species: Rat
    Sample Types: Tissue Homogenates
  21. Intestinal anti-inflammatory activity of Ground Cherry (Physalis angulata L.) standardized CO2 phytopharmaceutical preparation
    Authors: LD Almeida Ju, AEV Quaglio, CAR de Almeida, LC Di Stasi
    World J. Gastroenterol., 2017;23(24):4369-4380.
    Species: Rat
    Sample Types: Tissue Homogenates
  22. Carotid sinus nerve electrical stimulation in conscious rats attenuates systemic inflammation via chemoreceptor activation
    Authors: FM Santos-Alm, G Domingos-S, CA Meschiari, LC Fávaro, C Becari, JA Castania, A Lopes, TM Cunha, DJA Moraes, FQ Cunha, L Ulloa, A Kanashiro, GCSV Tezini, HC Salgado
    Sci Rep, 2017;7(1):6265.
    Species: Rat
    Sample Types: Plasma
  23. Impact of exercise training associated to pyridostigmine treatment on autonomic function and inflammatory profile after myocardial infarction in rats
    Authors: Bruno Rodrigues
    Int. J. Cardiol., 2016;0(0):.
    Species: Rat
    Sample Types: Plasma
  24. Correlation of Gut Microbiota Composition with Resistance to Experimental Autoimmune Encephalomyelitis in Rats
    Front Microbiol, 2016;7(0):2005.
    Species: Rat
    Sample Types: Cell Culture Supernates
  25. Effects of freshwater clam extract supplementation on time to exhaustion, muscle damage, pro/anti-inflammatory cytokines, and liver injury in rats after exhaustive exercise.
    Authors: Huang K, Wu W, Yang F, Chiu Y, Peng T, Hsu B, Liao K, Lee R
    Molecules, 2013;18(4):3825-38.
    Species: Rat
    Sample Types: Plasma
  26. Oligofructose supplementation (10%) during pregnancy and lactation does not change the inflammatory effect of concurrent trans fatty acid ingestion on 21-day-old offspring.
    Authors: Hachul A, Mennitti L, de Oliveira J, Moreno M, Okuda M, Dos Santos B, Oyama L, Ribeiro E, do Nascimento C, Pisani L
    Lipids Health Dis, 2013;12(0):59.
    Species: Rat
    Sample Types: Tissue Homogenates
  27. Depot-specific modulation of adipokine levels in rat adipose tissue by diet-induced obesity: the effect of aerobic training and energy restriction.
    Authors: Yamashita AS, Lira FS, Rosa JC, Paulino EC, Brum PC, Negrao CE, dos Santos RV, Batista ML, do Nascimento CO, Oyama LM, Seelaender M
    Cytokine, 2010;52(3):168-74.
    Species: Rat
    Sample Types: Tissue Homogenates
  28. Comparative studies on the interplay of testosterone, estrogen and progesterone in collagen induced arthritis in rats.
    Authors: Ganesan K, Balachandran C, Manohar BM, Puvanakrishnan R
    Bone, 2008;43(4):758-65.
    Species: Rat
    Sample Types: Serum
  29. IL-10 inhibits vascular smooth muscle cell activation in vitro and in vivo.
    Authors: Mazighi M, Pelle A, Gonzalez W, Mtairag el M, Philippe M, Henin D, Michel JB, Feldman LJ
    Am. J. Physiol. Heart Circ. Physiol., 2004;287(2):H866-71.
    Species: Rat
    Sample Types: Plasma

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Rat IL-10 DuoSet ELISA
By Anonymous on 08/11/2020
Application: Sample Tested: Serum,Tissue Homogenates,Lung tissue,Spleen tissue