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IL-17 Kit ELISA

IL17 Reactivité: Humain Colorimetric Sandwich ELISA 80-6000 pg/mL Cell Culture Supernatant, Plasma, Serum
N° du produit ABIN625013
  • Antigène Voir toutes IL-17 (IL17) Kits ELISA
    IL-17 (IL17) (Interleukin 17 (IL17))
    Reactivité
    • 7
    • 7
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Humain
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    80-6000 pg/mL
    Seuil minimal de détection
    80 pg/mL
    Application
    ELISA
    Fonction
    Human IL-17A ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Type d'échantillon
    Cell Culture Supernatant, Serum, Plasma
    Analytical Method
    Quantitative
    Specificité
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Réactivité croisée (Details)
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Sensibilité
    < 80 pg/mL
    Attributs du produit
    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data
    Ingrédients
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Stop Solution
    • Assay Diluent(s)
    • Lyophilized Standard
    • Biotinylated Detection Antibody
    • Streptavidin-Conjugated HRP
    • TMB One-Step Substrate
    Matériel non inclus
    • Distilled or deionized water
    • Precision pipettes to deliver 2 μL to 1 μL volumes
    • Adjustable 1-25 μL pipettes for reagent preparation
    • 100 μL and 1 liter graduated cylinders
    • Tubes to prepare standard and sample dilutions
    • Absorbent paper
    • Microplate reader capable of measuring absorbance at 450nm
    • Log-log graph paper or computer and software for ELISA data analysis
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  • Indications d'application
    Recommended Dilution for serum and plasma samples2 fold
    Volume d'échantillon
    100 μL
    Plaque
    Pre-coated
    Protocole
    1. Prepare all reagents, samples and standards as instructed in the manual.
    2. Add 100 μL of standard or sample to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 μL of prepared biotin antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 μL of prepared Streptavidin solution to each well.
    7. Incubate 45 min at RT.
    8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 μL of Stop Solution to each well.
    11. Read at 450 nm immediately.
    Préparation des réactifs
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
      2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
      3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 100 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 60 µL IL-17 standard from the vial of Item C, into a tube with 940 µL Assay Diluent A or 1x Assay Diluent B to prepare a 6,000 pg/mL standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 300 µL 300myl 300 µL 300 µL 300 µL 300 µL 60 µL standard + 940 µL 6,000 3,000 1,500 750 375 187.5 93.75 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
      5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
      6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 120-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 100 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a final 120 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    Procédure de l'essai
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
      2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
      3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
      4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
      5. Discard the solution. Repeat the wash as in step
      6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
      7. Discard the solution. Repeat the wash as in step
      8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
      9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
    Calcul des résultats

    Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
    Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Human IL-17 concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.1 1 10 Assay Diluent B Human IL-17 concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.1 1 10 Assay Diluent A
    Sensitivity: The minimum detectable dose of IL-17 is typically less than 80 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human IL-17 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 94.15 83-103 Plasma 92.34 82-104 Cell culture media 93.65 84-103
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93 94 91 Range ( %) 83-103 84-103 82-102 1:4 Average % of Expected 94 92 97 Range ( %) 82-103 83-102 85-104
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Précision du teste
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Conseil sur la manipulation
    Avoid repeated freeze-thaw cycles.
    Stock
    -20 °C
    Stockage commentaire
    The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
    Date de péremption
    6 months
  • Petek-Balc?, Çoban, Shugaiv, Türko?lu, Ulusoy, ?çöz, Pehlivan, Tüzün, Akman-Demir, Kürtüncü, Eraksoy: "Predictive value of early serum cytokine changes on long-term interferon beta-1a efficacy in multiple sclerosis." dans: The International journal of neuroscience, Vol. 125, Issue 5, pp. 352-6, (2015) (PubMed).

    Talaat, Mohamed, Bassyouni, Raouf: "Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus (SLE) patients: Correlation with disease activity." dans: Cytokine, Vol. 72, Issue 2, pp. 146-53, (2015) (PubMed).

    Chitrapriya, Rao, Lavu: "Interleukin-17 and interleukin-18 levels in different stages of inflammatory periodontal disease." dans: Journal of Indian Society of Periodontology, Vol. 19, Issue 1, pp. 14-7, (2015) (PubMed).

    Talaat, Sidek, Mosalem, Kholief: "Effect of bisphosphonates treatment on cytokine imbalance between TH17 and Treg in osteoporosis." dans: Inflammopharmacology, Vol. 23, Issue 2-3, pp. 119-25, (2015) (PubMed).

    Yi, Yu, Zhang, Song, Jiang, Du, Wang: "Cathelicidin-BF suppresses intestinal inflammation by inhibiting the nuclear factor-κB signaling pathway and enhancing the phagocytosis of immune cells via STAT-1 in weanling piglets." dans: International immunopharmacology, Vol. 28, Issue 1, pp. 61-9, (2015) (PubMed).

    Liu, Liao, Zhao, Wu, Yung, Chan, Yoshimura, Lu: "Increased expression of TLR2 in CD4(+) T cells from SLE patients enhances immune reactivity and promotes IL-17 expression through histone modifications." dans: European journal of immunology, Vol. 45, Issue 9, pp. 2683-93, (2015) (PubMed).

    Attia, Abdallah, El-Khateeb, Saad, Lotfi, Abdallah, El-Shennawy: "Serum Th17 cytokines in leprosy: correlation with circulating CD4(+) CD25 (high)FoxP3 (+) T-regs cells, as well as down regulatory cytokines." dans: Archives of dermatological research, Vol. 306, Issue 9, pp. 793-801, (2014) (PubMed).

    Ganjalikhani Hakemi, Ghaedi, Homayouni, Andalib, Hosseini, Rezaei: "Positive and Negative Regulation of Th17 Cell Differentiation: Evaluating The Impact of RORC2." dans: Cell journal, Vol. 16, Issue 3, pp. 343-52, (2014) (PubMed).

    Akelma, Cizmeci, Kanburoglu, Bozkaya, Catal, Mete, Kutukoglu, Namuslu: "Elevated level of serum osteopontin in school-age children with asthma." dans: Allergologia et immunopathologia, Vol. 42, Issue 4, pp. 275-81, (2014) (PubMed).

    Gao, Ding, Xiao, Li, Chen, Zhou, Wang, Wu, Shi: "Anti-inflammatory and anti-apoptotic effect of combined treatment with methylprednisolone and amniotic membrane mesenchymal stem cells after spinal cord injury in rats." dans: Neurochemical research, Vol. 39, Issue 8, pp. 1544-52, (2014) (PubMed).

    Arranz-Valsero, Schulze, Contreras-Ruiz, García-Posadas, López-García, Paulsen, Diebold: "Involvement of corneal epithelial cells in the Th17 response in an in vitro bacterial inflammation model." dans: Molecular vision, Vol. 19, pp. 85-99, (2013) (PubMed).

    Esendagli, Kurne, Sayat, Kilic, Guc, Karabudak: "Evaluation of Th17-related cytokines and receptors in multiple sclerosis patients under interferon ?-1 therapy." dans: Journal of neuroimmunology, Vol. 255, Issue 1-2, pp. 81-4, (2013) (PubMed).

    Karadag, Ertugrul, Bilgili, Takci, Akin, Calka: "Immunoregulatory effects of isotretinoin in patients with acne." dans: The British journal of dermatology, Vol. 167, Issue 2, pp. 433-5, (2012) (PubMed).

    Li, Jiang, Zhang, Yin, Ma, He, Shen: "Methotrexate attenuates the Th17/IL-17 levels in peripheral blood mononuclear cells from healthy individuals and RA patients." dans: Rheumatology international, Vol. 32, Issue 8, pp. 2415-22, (2012) (PubMed).

    Ulusoy, Tüzün, Kürtüncü, Türkoğlu, Akman-Demir, Eraksoy: "Comparison of the cytokine profiles of patients with neuronal-antibody-associated central nervous system disorders." dans: The International journal of neuroscience, Vol. 122, Issue 6, pp. 284-9, (2012) (PubMed).

    Wolfram, Rabensteiner, Grundtman, Böck, Mayerl, Parson, Almanzar, Hasenöhrl, Piza-Katzer, Wick: "T regulatory cells and TH17 cells in peri-silicone implant capsular fibrosis." dans: Plastic and reconstructive surgery, Vol. 129, Issue 2, pp. 327e-337e, (2012) (PubMed).

    Sibilano, Frossi, Calvaruso, Danelli, Betto, DallAgnese, Tripodo, Colombo, Pucillo, Gri: "The aryl hydrocarbon receptor modulates acute and late mast cell responses." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 189, Issue 1, pp. 120-7, (2012) (PubMed).

    Kouam, Kusari, Lamshöft, Tatuedom, Spiteller: "Sapelenins G-J, acyclic triterpenoids with strong anti-inflammatory activities from the bark of the Cameroonian medicinal plant Entandrophragma cylindricum." dans: Phytochemistry, Vol. 83, pp. 79-86, (2012) (PubMed).

    Ganjalikhani Hakemi, Ghaedi, Andalib, Hosseini, Rezaei: "Optimization of human Th17 cell differentiation in vitro: evaluating different polarizing factors." dans: In vitro cellular & developmental biology. Animal, Vol. 47, Issue 8, pp. 581-92, (2012) (PubMed).

    Kürtüncü, Tüzün, Türkoğlu, Petek-Balcı, Içöz, Pehlivan, Birişik, Ulusoy, Shugaiv, Akman-Demir, Eraksoy: "Effect of short-term interferon-β treatment on cytokines in multiple sclerosis: significant modulation of IL-17 and IL-23." dans: Cytokine, Vol. 59, Issue 2, pp. 400-2, (2012) (PubMed).

  • Antigène Voir toutes IL-17 (IL17) Kits ELISA
    IL-17 (IL17) (Interleukin 17 (IL17))
    Autre désignation
    IL-17 (IL17 Produits)
    Synonymes
    CTLA8 Kit ELISA, IL-17 Kit ELISA, IL-17A Kit ELISA, IL17 Kit ELISA, Ctla-8 Kit ELISA, Ctla8 Kit ELISA, Il17 Kit ELISA, ChIL-17 Kit ELISA, IL-17F Kit ELISA, IL17A Kit ELISA, CTLA-8 Kit ELISA, interleukin 17A Kit ELISA, IL17A Kit ELISA, Il17a Kit ELISA
    Sujet
    IL-17 enhances expression of the intracellular adhesion molecule-1 (ICAM-1) in human fibroblasts. Human IL-17 also stimulates epithelial, endothelial, or fibroblastic cells to secrete IL-6, IL-8, and G-CSF and prostaglandin E2. It functions as a mediator of angiogenesis that stimulates vascular endothelial cell migration and cord formation and regulates production of a variety of growth factors promoting angiogenesis (Numasaki et al). Mouse, rat, and human IL-17 can induce IL-6 secretion in mouse stromal cells, indicating that all homologs can recognize the mouse receptor. The Human IL-17 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human IL-17 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human IL-17 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-17 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human IL-17 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-17 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
    ID gène
    3605
    UniProt
    Q16552
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