EAE was induced in 8–12-week-old female Clec10a−/− and wild-type mice (C57BL/6) by s.c. immunization with 150 μg mouse MOG35–55 (MEVGWYRSPFSRVVHLYRNGK; Cambridge Research Biochemicals) in complete Freund’s adjuvant (CFA) supplemented with 4 mg/ml Mycobacterium tuberculosis (H37RA; Difco). Control animals were injected with a 1:1 PBS/CFA mixture. All animals received 200 ng pertussis toxin (Sigma) i.p. on days 0 and 2. Mice were examined daily for signs of EAE and scored as follows: 0, no clinical signs; 0.5, half limp tail; 1, complete limp tail; 1.5, lack of toe-spreading reflex; 2, half hind limb weakness; 2.5, hind limb weakness; 3, half hind limb paralysis; 3.5, incomplete hind limb paralysis; 4, complete hind limb paralysis; 4.5, diaphragmatic paralysis/paralysis of (one of the) front legs; and 5, death by EAE. At day 27, proliferation was determined in antigen-specific splenocytes and draining lymph node (DLN) cells by [3H]-thymidine incorporation following ex vivo restimulation with 25 μg/ml MOG35–55. Cytokine production was determined in supernatants following 72-h antigen restimulation by ELISA. The IL-17 ELISA kit was from R&D.
Il 17 elisa kit
Manufactured by R&D Systems
Sourced in United States
The IL-17 ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) used for the measurement of interleukin-17 (IL-17) levels in biological samples such as cell culture supernatants, serum, and plasma.
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Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Pertussis toxin, by Merck Group (1 mentions)
Il 11 elisa kit, by R&D Systems (1 mentions)
Atorvastatin, by Pfizer (1 mentions)
Human il 1β, by R&D Systems (1 mentions)
70 μm strainer, by SPL Life Sciences (1 mentions)
First strand cdna synthesis kit, by Takara Bio (1 mentions)
Hrp conjugated anti rabbit igg antibody, by Cell Signaling Technology (1 mentions)
Sybr premix ex taq, by Takara Bio (1 mentions)
Ethanol, by Merck Group (1 mentions)
6 protocols using il 17 elisa kit
1
Experimental Autoimmune Encephalomyelitis in Mice
2
Cytokine Quantification in Cell Culture
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Cell culture supernatants were assayed for IL-17, IFN–γ, and TGF–β. The concentration of IFN–γ in the supernatant was measured by sandwich ELISA (IFN-γ detection and biotinylated IFN-γ antibodies from BD Biosciences). The TGF–β concentration was measured with the standard Mv1Lu bioassay48 (link). The IL-17 concentration in the supernatant was measured using an IL-17 ELISA kit (R&D systems).
3
Atorvastatin Effects on Bone Metabolism
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Wister male rats (Hunan Slack Jingda Experimental Animals Co., Ltd.); Sumianxin, Suxingling (Institute of Military Veterinary Medicine, Academy of Military Medical Sciences); Atorvastatin (Pfizer); Enamel binders (Wuhan Hongji Dental Equipment Co., Ltd.); TRACP-5b ELISA kit, serum 25-hydroxyvitamin D3 ELISA kit, adrenal glucocorticoid ELISA kit, IL-1 ELISA kit, IL-11 ELISA kit as well as IL-17 ELISA kit (R&D); Human TRACP ELISA (Solarbio); RNAKL antibody, RANK antibody, OPG antibody, HMGCR antibody, β-actin antibody (Abcam).
4
Cytokine Quantification in Monocytes
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IL-17 levels in monocyte cell culture supernatants and in the control groups were determined by a IL-17 ELISA Kit (R&D system, USA). IL-6 and IL-33 levels were measured by ELISA kits (R&D system, USA). The detection threshold of the assay was found to be 1 ng/ml.
5
Cytokine Profiling of MDMs
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After MEHP treatment in vitro, cell-free supernatants from cultured MDMs were obtained to test the concentrations of cytokines by ELISA. Human IL-1β, IL-6, IL-8, TNF-α, IL-10, IL-12, IL-23, IFN-γ, IL-13, and IL-17 ELISA kits (R&D Systems, Minneapolis, MN, USA) were used according to the manufacturer’s instructions. Chemokine or cytokine concentrations were calculated based on linear regression standard curves, in which r2 was higher than 0.99.
6
Immunomodulatory Effects of Topical Imiquimod
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Ethanol (Merck, Germany), acetone (Duksan, Korea), hair removal cream (Veet; Reckitt Benckiser, France), IMQ (Dong-a Otsuka, Korea), Vaseline (Unilever Korea, Korea), 70 μm strainers (SPL life science, Korea), IL-17 ELISA kits (R&D Systems MN, USA), and PerCP-Cy5.5-Rat IgG1,κ (RTK2071, Biolegend, USA) were used. First Strand cDNA Synthesis Kits and SYBR Premix EX Taq were purchased from Takara Bio (Japan).
CD16/CD32, fluorescein isothiocyanate (FITC)-anti-mouse CD4 (Mouse CTL clone V4) and FITC-Rat IgG2b,κ (TNP-Keyhole Limpet Hemocyanin) were purchased from BD Biosciences (USA). FITC-anti-mouse γδ TCR (UC7-13D5) and phycoerythrin (PE)-anti-mouse IL-17A, FITC-Armenian Hamster IgG (eBio299Arm), PE-anti-mouse IL-17A (eBio17B7), PerCP-Cy5.5-anti-mouse IFNγ (XMG1.2), PE-anti-Rat IgG2a/κ (eBR2a) and TRIzol reagent were purchased from Thermo Fisher Scientific (USA). Rabbit anti-STAT1(D1K9Y), rabbit anti-phospho-STAT1 (Tyr701), rabbit anti-STAT3 (79D7), rabbit anti-phospho-STAT3 (D3A6), rabbit anti-STAT5 (D2O6Y), rabbit anti-phospho-STAT5 (C11C5) and HRP-conjugated anti-rabbit IgG antibodies were purchased from Cell Signaling (USA). Mouse anti-β-Actin (C4) antibodies and HRP-conjugated m-IgGκ BP were purchased from Santa Cruz Biotechnology (USA).
CD16/CD32, fluorescein isothiocyanate (FITC)-anti-mouse CD4 (Mouse CTL clone V4) and FITC-Rat IgG2b,κ (TNP-Keyhole Limpet Hemocyanin) were purchased from BD Biosciences (USA). FITC-anti-mouse γδ TCR (UC7-13D5) and phycoerythrin (PE)-anti-mouse IL-17A, FITC-Armenian Hamster IgG (eBio299Arm), PE-anti-mouse IL-17A (eBio17B7), PerCP-Cy5.5-anti-mouse IFNγ (XMG1.2), PE-anti-Rat IgG2a/κ (eBR2a) and TRIzol reagent were purchased from Thermo Fisher Scientific (USA). Rabbit anti-STAT1(D1K9Y), rabbit anti-phospho-STAT1 (Tyr701), rabbit anti-STAT3 (79D7), rabbit anti-phospho-STAT3 (D3A6), rabbit anti-STAT5 (D2O6Y), rabbit anti-phospho-STAT5 (C11C5) and HRP-conjugated anti-rabbit IgG antibodies were purchased from Cell Signaling (USA). Mouse anti-β-Actin (C4) antibodies and HRP-conjugated m-IgGκ BP were purchased from Santa Cruz Biotechnology (USA).
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