Rat IL-10 DuoSet ELISA Summary
* 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)
Scientific Data
Product Datasheets
Preparation and Storage
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).
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
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Citations: Showing 1 - 10
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Periodontitis and diabetes in pregnant rats: Maternal-fetal outcomes
Authors: Souza, SS;Lopes Cruz, L;Alves-Reis, AM;Costa, VQ;Moraes-Souza, RQ;Damasceno, DC;Volpato, GT;
Heliyon
Species: Rat
Sample Types: Serum
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Banisteriopsis caapi extract: Implications for Neuroinflammatory Pathways in Locus coeruleus Lesion Rodent Model
Authors: Graça, SC;Bustelli, IB;Santos, ÉVD;Fernandes, CG;Lanaro, R;Stilhano, RS;Linardi, A;Caetano, AL;
Journal of ethnopharmacology
Species: Rat
Sample Types: Tissue Homogenates
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Physical training attenuates systemic cytokine response and tissue damage triggered by apical periodontitis
Authors: Ferreira, RO;Pereira, MS;Souza-Monteiro, D;Frazão, DR;de Moura, JDM;Baia-da-Silva, DC;Bittencourt, LO;Balbinot, GS;Collares, FM;Lima, MLS;de Araújo, AA;Lima, RR;
Scientific reports
Species: Rat
Sample Types: Serum
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MANF protein expression is upregulated in immune cells in the ischemic human brain and systemic recombinant MANF delivery in rat ischemic stroke model demonstrates anti-inflammatory effects
Authors: Anttila, JE;Mattila, OS;Liew, HK;Mätlik, K;Mervaala, E;Lindholm, P;Lindahl, M;Lindsberg, PJ;Tseng, KY;Airavaara, M;
Acta neuropathologica communications
Species: Rat
Sample Types: Serum, Tissue Homogenates
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Analysis of anti-rheumatic activity of Nyctanthes arbor-tristis via in vivo and pharmacovigilance approaches
Authors: Sharma, A;Goel, A;Lin, Z;
Frontiers in pharmacology
Species: Rat
Sample Types: Serum
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Potential Benefits of Epidermal Growth Factor for Inhibiting Muscle Degrative Markers in Rats with Alcoholic Liver Damage
Authors: Xiao, Q;Chen, YH;Chen, YL;Chien, YS;Hsieh, LH;Shirakawa, H;Yang, SC;
International journal of molecular sciences
Species: Rat
Sample Types: Tissue Homogenates
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A nanofiber-hydrogel composite improves tissue repair in a rat model of Crohn's disease perianal fistulas
Authors: L Li, ZC Yao, A Parian, YH Yang, J Chao, J Yin, KJ Salimian, SK Reddy, A Zaheer, SL Gearhart, HQ Mao, FM Selaru
Science Advances, 2023-01-04;9(1):eade1067.
Species: Rat
Sample Types: Tissue Homogenates
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The superior beneficial effects of exercise training versus hormone replacement therapy on skeletal muscle of ovariectomized rats
Authors: SB Silva, K Honorato-S, SP Costa, TE Domingues, TMM da Cruz, CM Rodrigues, KB Costa, JM Dos Santos, VK da Silva L, TP Gaiad, AP Santos, MF Dias-Peixo, CC Coimbra, AM Dos Reis, RE Szawka, PHS Figueiredo, HS Costa, MX Oliveira, VA Mendonça, ACR Lacerda
Scientific Reports, 2022-05-24;12(1):8764.
Species: Rat
Sample Types: Tissue Homogenates
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Exposure to Nickel Oxide Nanoparticles Induces Acute and Chronic Inflammatory Responses in Rat Lungs and Perturbs the Lung Microbiome
Authors: MJ Jeong, S Jeon, HS Yu, WS Cho, S Lee, D Kang, Y Kim, YJ Kim, SY Kim
International Journal of Environmental Research and Public Health, 2022-01-04;19(1):.
Species: Rat
Sample Types: BALF
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The Prophylactic Effects of Glutamine on Muscle Protein Synthesis and Degradation in Rats with Ethanol-Induced Liver Damage
Authors: Q Xiao, YH Chen, SA Pratama, YL Chen, H Shirakawa, HC Peng, SC Yang
Nutrients, 2021-08-14;13(8):.
Species: Rat
Sample Types: Tissue Homogenates
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A novel rat model of chronic subdural hematoma: Induction of inflammation and angiogenesis in the subdural space mimicking human-like features of progressively expanding hematoma
Authors: X Xu, D Wang, Z Han, B Wang, W Gao, Y Fan, F Li, Z Zhou, C Gao, J Xiong, S Zhou, S Zhang, G Yang, R Jiang, J Zhang
Brain research bulletin, 2021-04-28;172(0):108-119.
Species: Rat
Sample Types: Tissue Homogenates
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Small intestine remodeling in male Goto-Kakizaki rats
Authors: JNB Pereira, GM Murata, FT Sato, AR Marosti, CRO Carvalho, R Curi
Physiological Reports, 2021-02-01;9(3):e14755.
Species: Rat
Sample Types: Tissue Homogenates
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The Poststroke Peripheral Immune Response Is Differentially Regulated by Leukemia Inhibitory Factor in Aged Male and Female Rodents
Authors: SM Davis, LA Collier, SJ Messmer, KR Pennypacke
Oxidative Medicine and Cellular Longevity, 2020-12-10;2020(0):8880244.
Species: Rat
Sample Types: Tissue Homogenates
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Impacts of fish oil on the gut microbiota of rats with alcoholic liver damage
Authors: YL Chen, H Shirakawa, NS Lu, HC Peng, Q Xiao, SC Yang
J. Nutr. Biochem., 2020-09-11;86(0):108491.
Species: Rat
Sample Types: Cell Culture Supernates
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GABA potentiate the immunoregulatory effects of Lactobacillus brevis BGZLS10-17 via ATG5-dependent autophagy in vitro
Authors: SS Baji?, J ?oki?, M Dini?, S Tomi?, N Popovi?, E Brdari?, N Goli?, M Tolina?ki
Sci Rep, 2020-01-28;10(1):1347.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2019-11-04;14(11):e0223800.
Species: Rat
Sample Types: Gingival Fluid
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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, 2019-10-27;8(11):.
Species: Rat
Sample Types: Tissue Lysates
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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, 2019-07-15;11(7):.
Species: Rat
Sample Types: Plasma
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Neuroinflammation in the NTS is associated with changes in cardiovascular reflexes during systemic inflammation
Authors: MR Amorim, JL de Deus, RA Cazuza, CMD Mota, LEV da Silva, GS Borges, ME Batalhão, EC Cárnio, LGS Branco
J Neuroinflammation, 2019-06-20;16(1):125.
Species: Rat
Sample Types: Plasma
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A diet including xanthan gum triggers a pro-inflammatory response in Wistar rats inoculated with Walker 256 cells
Authors: AB Silva Risc, NIP Neto, K Gascho, M Carnier, DA de Miranda, FP Silva, VT Boldarine, M Seelaender, EB Ribeiro, LM Oyama, CM Oller do N
PLoS ONE, 2019-06-18;14(6):e0218567.
Species: Rat
Sample Types: Tissue Homogenates
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Metformin Augments Anti-Inflammatory and Chondroprotective Properties of Mesenchymal Stem Cells in Experimental Osteoarthritis
Authors: MJ Park, SJ Moon, JA Baek, EJ Lee, KA Jung, EK Kim, DS Kim, JH Lee, SK Kwok, JK Min, SJ Kim, SH Park, ML Cho
J. Immunol., 2019-05-29;0(0):.
Species: Human, Rat
Sample Types: Cell Culture Supernates, Serum
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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, 2019-05-09;20(9):.
Species: Rat
Sample Types: Serum
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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, 2019-04-27;24(9):.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2019-02-21;9(1):2481.
Species: Rat
Sample Types: Plasma
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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, 2019-01-29;9(1):918.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2018-12-31;3(6):766-778.
Species: Rat
Sample Types: Serum
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Obesity and Type 2 Diabetes mellitus induce lipopolysaccharide tolerance in rat neutrophils
Authors: WMT Kuwabara, CNF Yokota, R Curi, TC Alba-Loure
Sci Rep, 2018-12-03;8(1):17534.
Species: Rat
Sample Types: BALF
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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, 2018-10-15;15(1):288.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2018-09-30;19(10):.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2018-07-21;23(7):.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2018-03-20;15(1):90.
Species: Rat
Sample Types: Cell Culture Supernates
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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, 2018-01-18;11(0):11-24.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2018-01-01;23(0):2515690X18765.
Species: Rat
Sample Types: Serum
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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., 2017-12-13;2017(0):5801768.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2017-07-24;7(1):6265.
Species: Rat
Sample Types: Plasma
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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., 2017-06-28;23(24):4369-4380.
Species: Rat
Sample Types: Tissue Homogenates
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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., 2017-05-01;96(5):1071-1084.
Species: Rat
Sample Types: Whole Tissue
Applications: ELISA Capture -
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, 2017-03-18;0(0):.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2017-02-12;643(0):27-31.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2017-01-20;7(0):40373.
Species: Rat
Sample Types: Tissue Homogenates
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Correlation of Gut Microbiota Composition with Resistance to Experimental Autoimmune Encephalomyelitis in Rats
Front Microbiol, 2016-12-15;7(0):2005.
Species: Rat
Sample Types: Cell Culture Supernates
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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., 2016-10-26;0(0):.
Species: Rat
Sample Types: Plasma
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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, 2013-05-01;12(0):59.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2013-03-26;18(4):3825-38.
Species: Rat
Sample Types: Plasma
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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, 2010-08-21;52(3):168-74.
Species: Rat
Sample Types: Tissue Homogenates
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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, 2008-06-10;43(4):758-65.
Species: Rat
Sample Types: Serum
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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., 2004-04-08;287(2):H866-71.
Species: Rat
Sample Types: Plasma
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