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Interleukin 6 Kit ELISA (IL6) Kit ELISA

IL6 Reactivité: Humain Colorimetric Sandwich ELISA 7.8-500 pg/mL Cell Culture Supernatant, Plasma, Serum, Tissue Homogenate, Urine
Pubmed (30) Independent Validation (1)
N° du produit ABIN365163
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  • Antigène
    Interleukin 6 (IL6)
    • 14
    • 9
    • 8
    • 5
    • 4
    • 4
    • 4
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    • 2
    • 2
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    Méthode de détection
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    7.8-500 pg/mL
    Seuil minimal de détection
    7.8 pg/mL
    For the quantitative determination of human interleukin 6 (IL-6) concentrations in serum, plasma, cell culture supernates, tissue homogenates and urine.
    Type d'échantillon
    Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate, Urine
    Analytical Method
    This assay has high sensitivity and excellent specificity for detection of human IL-6.
    Réactivité croisée (Details)
    Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between the target antigen and all analogues for other species. Therefore, cross reaction may still exist.
    2.453 pg/mL
    • Assay plate (12 × 8 coated Microwells)
    • Standard (freeze dried)
    • Biotin-antibody (100 × concentrate)
    • HRP-avidin (100 × concentrate)
    • Biotin-antibody Diluent
    • HRP-avidin Diluent
    • Sample Diluent
    • Wash Buffer (25 × concentrate)
    • TMB Substrate
    • Stop Solution
    • Adhesive Strip (for 96 wells)
    • Instruction manual
    Matériel non inclus
    • Microplate reader capable of measuring absorbance at 450nm, with the correction wavelength set at 540nm or 570nm.
    • An incubator which can provide stable incubation conditions up to 37°C ± 0.5°C.
    • Squirt bottle, manifold dispenser or automated microplate washer.
    • Absorbent paper for blotting the microtiter plate.
    • 100mL and 500mL graduated cylinders.
    • Deionized or distilled water.
    • Pipettes and pipette tips.
    • Test tubes for dilution.
    Discover our best selling IL6 Kit ELISA
  • Indications d'application
    • The supplier is only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
    • Samples to be used within 5 days may be stored at 2-8°C, otherwise samples must be stored at -20°C (≤ 1 month) or -80°C (≤ 2 months) to avoid loss of bioactivity and contamination.
    • Grossly hemolyzed samples are not suitable for use in this assay.
    • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
    • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
    • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts of certain chemicals.
    • Owing to the possibility of mismatching between antigens from another resource and antibodies used in this supplier's kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by this supplier's products.
    • Influenced by factors including cell viability, cell number and cell sampling time, samples from cell culture supernatant may not be recognized by the kit.
    • Fresh samples without long time storage are recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.

    Detection wavelength: 450 nm

    Information on standard material:
    Depending on the antigen to be detected, standards can be either native or recombinant protein. The recombinant proteins are being expressed in CHO cells in most cases. Please inquire for more information. The formulation of auxiliary material in the standard is considered proprietary information, however it does not contain any poisonous substance. Proclin 300 (1:3000) is used as preservative.

    Information on reagents:
    In most cases the stop solution provided is 1 N H2SO4. The formulation of wash solution is proprietary information. None of the components contain (sodium) azide, thimerosal, 2-mercaptoethanol (2-ME) or any other poisonous materials. For the sandwich method kits, the sample diluent, antibody diluent, enzyme diluent and standard all contain BSA.

    Information on antibodies:
    The antibodies provided in different kits vary in regards to clonality and host. Some antibodies are affinity purified, some are Protein A

    Volume d'échantillon
    100 μL
    Durée du test
    1 - 4.5 h
    This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for IL-6 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-6 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for IL-6 is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-6 bound in the initial step. The color development is stopped and the intensity of the color is measured.
    Préparation des réactifs
    • Biotin-antibody (1×) - Centrifuge the vial before opening.
      Biotin-antibody requires a 100-fold dilution. The suggested dilution is 10µL of Biotin-antibody + 990µL of Biotin-antibody Diluent.
    • HRP-avidin (1×) - Centrifuge the vial before opening.
      HRP-avidin requires a 100-fold dilution. The suggested dilution is 10µL of HRP-avidin + 990µL of HRP-avidin Diluent.
    • Wash Buffer (1×) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20mL of Wash Buffer Concentrate (25×) into deionized or distilled water to prepare 500mL of Wash Buffer (1×).
    • Standard - Centrifuge the standard vial at 6000-10000rpm for 30s.
      Reconstitute the Standard with 1ml of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.
      Pipette 250µL of Sample Diluent into each tube. Use the stock solution to produce a 2-fold dilution series. Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard. Sample Diluent serves as the zero standard (0ng/mL).
    • Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
    • Bring all reagents to room temperature (18-25°C) before use for 30 min.
    • Prepare fresh standard for each assay. Use within 4 hours and discard after use.
    • Making serial dilution in the wells directly is not permitted.
    • Please carefully reconstitute Standards according to the instruction. Avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µL when pipetting.
    • It is recommended to use distilled water to prepare reagents and samples. Using contaminated water or container for reagent preparation will influence detection result.
    Prélèvement de l'échantillon
    • Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4 °C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
    • Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2-8 °C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
    • Tissue Homogenates: Rinse 100 mg tissue with 1× PBS, homogenize in 1mL of 1× PBS and store overnight at -20 °C. After two freeze-thaw cycles to break the cell membranes, centrifuge the homogenates for 5 minutes at 5000 × g, 2-8 °C. Remove and assay the supernate immediately. Alternatively, aliquot and store samples at -20 °C or -80 °C. Centrifuge the sample again after thawing before the assay. Avoid repeated freeze-thaw cycles.
    Procédure de l'essai
    • 1. Prepare all reagents, working standards and samples as directed in the respective sections.
    • 2. Refer to the Assay Layout Sheet to determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.
    • 3. Add 100µL of standard or sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C. A plate layout is provided to record standards and samples assayed.
    • 4. Remove the liquid of each well, don't wash.
    • 5. Add 100µL of Biotin-antibody (1×) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C. (Biotin-antibody (1×) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)
    • 6. Aspirate each well and wash, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (200µL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher and let it stand for 2 minutes, complete removal of liquid at each step is essential for 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.
    • 7. Add 100µL of HRP-avidin (1×) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
    • 8. Repeat the aspiration/wash process for five times as in step 6.
    • 9. Add 90µL of TMB Substrate to each well. Incubate for 20 minutes at 37°C. Protect from light.
    • 10. Add 50µL of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
    • 11. Determine the optical density of each well within 5 minutes using a microplate reader set to 450nm. If wavelength correction is available, set to 540nm or 570nm. Subtract readings at 540nm or 570nm from the readings at 450nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450nm without correction may be higher and less accurate.
    • The experiment's final results will be closely related to validity of the products, operation skills of the end users and the environmental conditions.
    • Samples or reagents addition: Please use the freshly prepared Standard. Please carefully add samples to wells and mix gently to avoid foaming. Do not touch the well wall as possible. For each step in the procedure, total dispensing time for addition of reagents or samples to the assay plate should not exced 10 minutes. This will ensure equal elapsed time for each pipetting step, without interruption. Duplication of all standards and specimens, although not required, is recommended. To avoid cross-contamination, change pipette tips between reagent additions. Also, use separate reservoirs for each reagent.
    • Incubation: To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. Incubation time and temperature must be observed.
    • Washing: The wash procedure is critical. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting and remove any drop of water and fingerprint on the bottom of the plate. Insufficient washing will result in poor precision and falsely elevated absorbance reading. When using an automated plate washer, adding a 30 second soak period following the addition of wash buffer and/or rotating the plate 180 degrees between wash steps may improve assay precision.
    • Controlling of reaction time: Observe the change of color after adding TMB Substrate (e.g. observation once every 10 minutes), TMB Substrate should change from colorless or light blue to gradations of blue. If the color is too deep, add Stop Solution in advance to avoid excessively strong reaction which will result in inaccurate absorbance reading.
    • TMB Substrate is easily contaminated. TMB Substrate should remain colorless or light blue until added to the plate. Please protect it from light.
    • Stop Solution should be added to the plate in the same order as the TMB Substrate. The color developed in the wells will turn from blue to yellow upon addition of the Stop Solution. Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the TMB Substrate.
    Calcul des résultats

    Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.
    Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the target antigen concentration versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
    If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

    Précision du teste
    Intra-assay precision (precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess precision.
    Inter-assay precision (precision between assays): Three samples of known concentration were tested in twenty assays to assess precision.
    • Intra-assay: CV% less than 8%
    • Inter-assay: CV% less than 10%
    For Research Use only
  • Validation #028773 (ELISA)
    'Independent Validation' signe
    Alamo Laboratories Inc
    Numéro du lot
    Application validée
    Contrôle positif
    Human serum
    Contrôle négative
    Mouse brain lysate
    Signal was detected in positive control samples but not in negative control samples.
    'Independent Validation' signe
    Validation Images
    Anticorps primaire
    • Antigen: Interleukin 6 (IL-6)
    • Catalog number: ABIN365163
    • Batch number: Q03097538
    Anticorps secondaire
    Full Protocol
    • To each well, 100 µL of standard or sample were added. Plate was incubated for 2 h at 37°C. The
    • liquid from each well was aspirated but wells were not washed.
    • 100 µL of Biotin-antibody (1x) was added to each well and the plate was incubated for 1 h at 37°C.
    • The liquid from each well was aspirated and wells were washed three times with 200 µL of Wash Buffer
    • (1X) each time x 2 min.
    • 100 µL of HRP-avidin (1x) was added to each well and the plate was incubated for 1 h at 37°C.
    • The liquid from each well was aspirated and wells were washed five times with 200 µL of Wash Buffer
    • (1X) each time x 2 min.
    • 90 µL of TMB Substrate was added to each well and the plate was incubated for 25 min at 37°C.
    • 50 µL of Stop Solution to each well and the contents were mixed by tapping gently. Absorbance of each
    • well was measured at 450 nm and 540 nm within 5 min using a microplate reader.
    • The readings at 540 nm were subtracted from those at 450 nm to correct for optical imperfections in the
    • plate.
    • The triplicate readings for each sample were averaged and the average zero standard optical density
    • subtracted. The corrected average-value was tabulated as Average Absorbance. A standard curve was
    • generated by plotting the mean OD value for each standard on the X-axis against the concentration on
    • the Y-axis using Excel. Standard curve was generated by regression analysis with four parameter
    • logistic.
    • An equation (y = 248.75x4-542.17x3+396.6x2+131.48x+2.0519) was derived from the standard
    • curve and used to calculate IL-6 concentrations based on their Average Absorbance values.
  • Précaution d'utilisation
    The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
    Conseil sur la manipulation
    • The kit should not be used beyond the expiration date on the kit label.
    • Do not mix or substitute reagents with those from other lots or sources.
    • If samples generate values higher than the highest standard, dilute the samples with Sample Diluent and repeat the assay.
    • Any variation in Sample Diluent, operator, pipetting technique, washing technique, incubation time/temperature and kit age can cause variation in binding.
    • This assay is designed to eliminate interference by soluble receptors, binding proteins and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
    4 °C/-20 °C
    Stockage commentaire
    May be stored at 2-8°C for up to 1 month. For long term storage, please store at -20°C. Try to keep assay plate in a sealed aluminium foil bag and avoid dampness.
    Date de péremption
    6 months
  • Yang, Yu, Dong, Zhang, Du, Zhu, Che, Wang, Shen, Jiang: "Serum macrophage migration inhibitory factor concentrations correlate with prognosis of traumatic brain injury." dans: Clinica chimica acta; international journal of clinical chemistry, Vol. 469, pp. 99-104, 2018 (PubMed).

    Yu, Huang, Po, Tan, Wang, Zhou, Meng, Yuan, Zhou, Li, Wang, Wang, Jiang: "Low-Level Tragus Stimulation for the Treatment of Ischemia and Reperfusion Injury in Patients With ST-Segment Elevation Myocardial Infarction: A Proof-of-Concept Study." dans: JACC. Cardiovascular interventions, Vol. 10, Issue 15, pp. 1511-1520, 2018 (PubMed).

    Chen, Cheng, Shao, Shentu, Fu: "Macrophage migration inhibitory factor as a serum prognostic marker in patients with aneurysmal subarachnoid hemorrhage." dans: Clinica chimica acta; international journal of clinical chemistry, Vol. 473, pp. 60-64, 2018 (PubMed).

    Wang, Yan, Peng, Jiang, He, Peng, Chen, Ye, Zhuo: "Functional Role of SUV39H1 in Human Renal Tubular Epithelial Cells Under High-glucose Ambiance." dans: Inflammation, Vol. 41, Issue 1, pp. 1-10, 2018 (PubMed).

    Golkhalkhali, Rajandram, Paliany, Ho, Wan Ishak, Johari, Chin: "Strain-specific probiotic (microbial cell preparation) and omega-3 fatty acid in modulating quality of life and inflammatory markers in colorectal cancer patients: a randomized controlled trial." dans: Asia-Pacific journal of clinical oncology, Vol. 14, Issue 3, pp. 179-191, 2018 (PubMed).

    Liu, Liu, Feng, Xiao: "Kinin B1 receptor as a novel, prognostic progression biomarker for carotid atherosclerotic plaques." dans: Molecular medicine reports, Vol. 16, Issue 6, pp. 8930-8936, 2018 (PubMed).

    Weeratunga, Herath, Kim, Lee, Kim, Lee, Lee, Chathuranga, Chathuranga, Yang, Ma, Lee: "Dense Granule Protein-7 (GRA-7) of Toxoplasma gondii inhibits viral replication in vitro and in vivo." dans: Journal of microbiology (Seoul, Korea), Vol. 55, Issue 11, pp. 909-917, 2018 (PubMed).

    Zhu, Jia, Luo, Wang: "Polyriboinosinic-polyribocytidylic acid facilitates interleukin-6, and interleukin-8 secretion in human dermal fibroblasts via the JAK/STAT3 and p38 MAPK signal transduction pathways." dans: Cytokine, Vol. 102, pp. 1-6, 2018 (PubMed).

    Bi, Wang, Wang, Sun, Dong, Meng, Li: "Selenium inhibits Staphylococcus aureus-induced inflammation by suppressing the activation of the NF-κB and MAPK signalling pathways in RAW264.7 macrophages." dans: European journal of pharmacology, Vol. 780, pp. 159-65, 2017 (PubMed).

    Guan, Chen, Zuo, Guo, Peng, Wang, Yin, Li: "Contrast Media-Induced Renal Inflammation Is Mediated Through HMGB1 and Its Receptors in Human Tubular Cells." dans: DNA and cell biology, Vol. 36, Issue 1, pp. 67-76, 2017 (PubMed).

    Yu, Li, Deng, Yu, Luo, Sun: "Synovial fluid concentrations of cold-inducible RNA-binding protein are associated with severity in knee osteoarthritis." dans: Clinica chimica acta; international journal of clinical chemistry, Vol. 464, pp. 44-49, 2017 (PubMed).

    Mao, Cao, Wang, Wang, Chen, Wang, Xing, Ren, Lv, Dong, Chen, Chen, Wang, Yan: "The Salutary Influence of Forest Bathing on Elderly Patients with Chronic Heart Failure." dans: International journal of environmental research and public health, Vol. 14, Issue 4, 2017 (PubMed).

    Xiao, Gong, Chen, Yu, Wang, Zhang, Dou, Liu, Cheng, Lu, Yuan, Li, Zhao: "IL-6 promotes epithelial-to-mesenchymal transition of human peritoneal mesothelial cells possibly through the JAK2/STAT3 signaling pathway." dans: American journal of physiology. Renal physiology, Vol. 313, Issue 2, pp. F310-F318, 2017 (PubMed).

    Li, Qiu, Li, Chen, Zhu, Chai: "Assessment of the hemolysis and endothelial cell cytotoxicity induced by residual linear alkylbenzene sulfonates on pharmaceutical rubber stoppers based on HPLC-ESI-MS." dans: Biomedical chromatography : BMC, 2015 (PubMed).

    Ambreen, Ismail, Qureshi: "Association of gene polymorphism with serum levels of inflammatory and angiogenic factors in Pakistani patients with age-related macular degeneration." dans: Molecular vision, Vol. 21, pp. 985-99, 2015 (PubMed).

    Fortis-Barrera, Alarcón-Aguilar, Banderas-Dorantes, Díaz-Flores, Román-Ramos, Cruz, García-Macedo: "Cucurbita ficifolia Bouché (Cucurbitaceae) and D-chiro-inositol modulate the redox state and inflammation in 3T3-L1 adipocytes." dans: The Journal of pharmacy and pharmacology, Vol. 65, Issue 10, pp. 1563-76, 2014 (PubMed).

    Guo, Jiang, Tang, Si, Jiao: "The association of serum vascular endothelial growth factor and ferritin in diabetic microvascular disease." dans: Diabetes technology & therapeutics, Vol. 16, Issue 4, pp. 224-34, 2014 (PubMed).

    Chen, Xu, Chen, Chen, Zhang, Ren, Xu: "Effect of oridonin-mediated hallmark changes on inflammatory pathways in human pancreatic cancer (BxPC-3) cells." dans: World journal of gastroenterology : WJG, Vol. 20, Issue 40, pp. 14895-903, 2014 (PubMed).

    Cheng, Zhong, Xiang, Zeng, Cai, Zhao: "Soluble receptor for advanced glycation end products in critically ill patients and its associations with other clinical markers and 28-day mortality." dans: Clinical interventions in aging, Vol. 9, pp. 1981-6, 2014 (PubMed).

    Qu, Wang, Deng, Wei, Deng: "Associations between longer habitual day napping and non-alcoholic fatty liver disease in an elderly Chinese population." dans: PLoS ONE, Vol. 9, Issue 8, pp. e105583, 2014 (PubMed).

  • Antigène
    Interleukin 6 (IL6)
    Autre désignation
    Interleukin 6 (IL-6) (IL6 ELISA Kit Extrait)
    BSF2, HGF, HSF, IFNB2, IL-6, Il-6, ILg6, Ifnb2, il6, CHIL-6, interleukin 6, interleukin-6, IL6, Il6, il-6, IL-6
    Synonyms: BSF2, HGF, HSF, IFNB2, IL-6, B cell stimulatory factor-2|B-cell differentiation factor|CTL differentiation factor|OTTHUMP00000158544|hybridoma growth factor|interleukin 6|interleukin BSF-2
    Signalisation TLR, Hormone Transport, Negative Regulation of Hormone Secretion, Myometrial Relaxation and Contraction, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Regulation of Carbohydrate Metabolic Process, Autophagy, Cell RedoxHomeostasis, Cancer Immune Checkpoints, Inflammasome
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