Human IL-6(Interleukin 6) ELISA Kit

    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H0102

      Synonyms:IL6, BSF2, HGF, HSF, IFNB2, CDF

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Human

      Detection Range: 7.81~500 pg/mL

      Sensitivity: 4.69 pg/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human IL-6 concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IL-6. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IL-6 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IL-6, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human IL-6. You can calculate the concentration of Human IL-6 in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 7.81—500 pg/mL
      Sensitivity 4.69 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human IL-6 in samples. No significant cross-reactivity or interference between Human IL-6 and analogues was observed.

      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration
      (pg/mL)
      O.D Average Corrected
      500 2.463
      2.487
      2.475 2.407
      250 1.612
      1.636
      1.624 1.556
      125 1.012
      0.978
      0.995 0.927
      62.5 0.513
      0.533
      0.523 0.455
      31.25 0.257
      0.245
      0.251 0.183
      15.63 0.174
      0.17
      0.172 0.104
      7.81 0.119
      0.123
      0.121 0.053
      0 0.067
      0.069
      0.068 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL-6 were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IL-6 were tested on 3 different plates, 20 replicates in each plate.

      Intra-assay Precision Inter-assay Precision
      Sample 1 2 3 1 2 3
      n 20 20 20 20 20 20
      Mean (pg/mL) 25.30 67.60 210.10 24.20 66.50 197.40
      Standard deviation 1.70 2.80 7.80 1.60 3.40 6.70
      C V (%) 6.72 4.14 3.71 6.61 5.11 3.39

      Recovery

      The recovery of Human IL-6 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

      Sample Type Range (%) Average Recovery (%)
      Serum (n=5) 97-110 102
      EDTA plasma (n=5) 92-108 99
      Cell culture media (n=5) 95-109 101

      Linearity

      Samples were spiked with high concentrations of Human IL-6 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 100-111 89-103 88-101
      Average (%) 105 97 93
      1:4 Range (%) 97-108 83-96 94-109
      Average (%) 102 90 100
      1:8 Range (%) 94-109 83-97 95-110
      Average (%) 101 89 102
      1:16 Range (%) 99-112 84-96 93-105
      Average (%) 106 88 99

      Kit components & Storage

      An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 6 months
      Biotinylated Detection Ab Diluent 1 vial, 14 mL
      HRP Conjugate Diluent 1 vial, 14 mL
      Concentrated Wash Buffer (25×) 1 vial, 30 mL
      Substrate Reagent 1 vial, 10 mL 4℃(shading light)
      Stop Solution 1 vial, 10 mL 4℃
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37℃
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      elisa assay procedure 1

      1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.

      elisa assay procedure 2

      2.Remove the liquid.

      elisa assay procedure 3

      3.Add 100 μL Biotinylated Detection Ab. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.

      elisa assay procedure 4

      4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.

      elisa assay procedure 5

      5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

      elisa assay procedure 6

      6.Add 50 μL Stop Solution.

      elisa assay procedure 7

      7.Read at 450 nm immediately. Calculation of results.

      Citations

      1. Publication: Liao Y, Liu P, Guo F, et al. Oxidative Burst Of Circulating Neutrophils Following Traumatic Brain Injury In Human[J]. Plos One, 2013.
        Sample Type: Plasma
      2. Publication: Tang J N, Shen L, Liu C L, et al. Plasma Levels Of C1q/TNF-Related Protein 1 And Interleukin 6 In Patients With Acute Coronary Syndrome Or Stable Angina Pectoris[J]. American Journal of The Medical Sciences, 2015, 349(2): 130-136.
        Sample Type: Plasma
      3. Publication: Cheng P, Hua W, Zhang W J, et al. Inhibitory Effect Of miR-125b On Hepatitis C Virus Core Protein-Induced TLR2/MyD88 Signaling In THP-1 Macrophages[J]. World Journal of Gastroenterology, 2016, 22(17): 4354-4361.
        Sample Type: Culture supernatant
      4. Publication: Tian P, Li B, He C, et al. Antidiabetic (Type 2) Effects Of Lactobacillus G15 And Q14 In Rats Through Regulation Of Intestinal Permeability And Microbiota[J]. Food & Function, 2016.
        Sample Type: Plasma and Serum
      5. Publication: Zhao Y, Jiang C, He J, et al. Multifunctional Dextran Sulfate-Coated Reconstituted High Density Lipoproteins Target Macrophages And Promote Beneficial Antiatherosclerotic Mechanisms[J]. Bioconjugate Chemistry, 2016.
        Sample Type: Culture supernatant
      6. Publication: Li W, Dong H, Huang Y, et al. Clonorchis Sinensis Co-Infection Could Affect The Disease State And Treatment Response Of Hbv Patients[J]. Plos Neglected Tropical Diseases, 2016.
        Sample Type: Culture supernatants and Serum
      7. Publication: Veena V K, Popavath R N, Kennedy K, et al. In Vitro Antiproliferative, Pro-Apoptotic, Antimetastatic And Anti-Inflammatory Potential Of 2, 4-Diacteylphloroglucinol (DAPG) By Pseudomonas Aeruginosa Strain FP10[J]. Apoptosis, 2015, 20(10): 1281-1295.
        Sample Type: Culture supernatant
      8. Publication: Hao S Y, Ji J J, Zhao H T,et al. Mitochondrion-Targeted Peptide SS-31 Inhibited Oxidized Low-Density Lipoproteins-Induced Foam Cell Formation Through Both ROS Scavenging And Inhibition Of Cholesterol Influx In Raw264.7 Cells[J]. Molecules, 2015, 20(12): 21287-21297.
        Sample Type: Cell lysis
      9. Publication: Li L, Zhang J, Zhao M, et al. Applied Value Of Monitoring Serum Hepcidin In Differential Diagnosis Of Infection Versus Tumor Fevers[J]. Journal of Huazhong University of Science and Technology-Medical Sciences, 2017, 37(2): 253-256.
        Sample Type: Plasma
      10. Publication: He W, Ren Y, Wang X, et al. C Reactive Protein And Enzymatically Modified LDL Cooperatively Promote Dendritic Cell-Mediated T Cell Activation[J]. Cardiovascular Pathology, 2017, 29: 1-6.
        Sample Type: Culture supernatant
      11. Publication: Chen W H, Luo G F, Lei Q, et al. Overcoming The Heat Endurance Of Tumor Cells By Interfering The Anaerobic Glycolysis Metabolism For Improved Photothermal Therapy[J]. Acs Nano, 2017, 11(2): 1419-1431.
        Sample Type: Culture supernatant
      12. Publication: Lei C T, Su H, Ye C, et al. The Classic Signalling And Trans-signalling Of Interleukin-6 Are Both Injurious In Podocyte Under High Glucose Exposure[J]. Journal of Cellular and Molecular Medicine, 2017.
        Sample Type: Serum
      13. Publication: Xu H, Sun Q, Lu L, et al. MicroRNA-218 Acts By Repressing TNFR1-Mediated Activation Of NF-κB, Which Is Involved In MUC5AC Hyper-Production And Inflammation In Smoking-Induced Bronchiolitis Of COPD[J]. Toxicology Letters, 2017, 280: 171-180.
        Sample Type: Serum and culture supernatant
      14. Publication: Cao Z, Chen L. Inhibition Of miR27a Suppresses The Inflammatory Response Via The p38/MAPK Pathway In Intervertebral Disc Cells[J]. Experimental and Therapeutic Medicine, 2017, 14(5): 4572-4578.
        Sample Type: Culture supernatant
      15. Publication: Li Z, Tang Y, Tang N, et al. High Anti-Human Cytomegalovirus Antibody Levels Are Associated With The Progression Of Essential Hypertension And Target Organ Damage In Han Chinese Population[J]. Plos One, 2017.
        Sample Type: Serum
      16. Publication: Wang N N, Guo J, Liu F D, et al. Depot-Specific Inflammation With Decreased Expression Of ATM2 In White Adipose Tissues Induced By High-Margarine/Lard Intake[J]. Plos One, 2017.
        Sample Type: Culture supernatant
      17. Publication: Liu Q, Wu J B, Song J Q, et al. Particulate Matter 2.5 Regulates Lipid Synthesis And Inflammatory Cytokine Production In Human SZ95 Sebocytes[J]. International Journal of Molecular Medicine, 2017, 40(4): 1029-1036.
        Sample Type: Culture supernatant
      18. Publication: Pan Y Y, Zhou F H, Song Z H, et al. Oleanolic acid protects against pathogenesis of atherosclerosis, possibly via FXR-mediated angiotensin (Ang)-(1-7) upregulation[J]. Biomedicine & Pharmacotherapy, 2018, 97: 1694-1700.
        Sample Type: Serum and culture supernatant
      19. Publication: Lu A Q, Wang Z R, Wang S C. Role Of miR-589-3p In Human Lumbar Disc Degeneration And Its Potential Mechanism[J]. Experimental and Therapeutic Medicine, 2018, 15: 1616-1621.
        Sample Type: Culture supernatant
      20. Publication: Jiang Z, Wang W H, Guo C C. Tetrahydroxy Stilbene Glucoside Ameliorates H2O2-Induced Human Brain Microvascular Endothelial Cell Dysfunction In Vitro By Inhibiting Oxidative Stress And Inflammatory Responses[J]. Molecular Medicine Reports, 2017, 16(4): 5219-5224.
        Sample Type: Culture supernatant
      21. Publication: Liu J, Lu C Y, Zhou H, et al. Flexible Gold Electrode Array For Multiplexed Immunoelectrochemical Measurement Of Three Protein Biomarkers For Prostate Cancer[J]. ACS Applied Materials & Interfaces, 2014, 6(22): 20137-20143.
        Sample Type: /
      22. Publication: Zhao J, Liang Y, Song F, et al. TSG Attenuates LPC-Induced Endothelial Cells Inflammatory Damage Through Notch Signaling Inhibition[J]. IUBMB Life, 2016, 68(1): 37-50.
        Sample Type: Culture supernatant
      23. Publication: Park K S, Kim S H, Das A, et al. TLR3-/4-Priming Differentially Promotes Ca2+Signaling And Cytokine Expression And Ca2+-Dependently Augments Cytokine Release In hMSCs[J]. Scientific Reports, 2016.
        Sample Type: Culture supernatant
      24. Publication: Kruger M J, Conradie R, Conradie M, et al. ADSC-conditioned media elicit an ex vivo anti-inflammatory macrophage response[J]. Journal of molecular endocrinology, 2018: JME-18-0078.
        Sample Type: Serumandconditioned media
      25. Publication: Akash M S H, Rehman K, Liaqat A, et al. Biochemical investigation of gender-specific association between insulin resistance and inflammatory biomarkers in types 2 diabetic patients[J]. Biomedicine & Pharmacotherapy, 2018, 106: 285-291.
        Sample Type: Serum
      26. Publication: Wang F, Li X, Huang L, et al. High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura[J]. Pediatric Allergy, Immunology, and Pulmonology, 2018, 31(2): 66-72.
        Sample Type: Serum
      27. Publication: Zhou P, Liu H, Wu Y, et al. Propofol Promotes Ankle Fracture Healing in Children by Inhibiting Inflammatory Response[J]. Medical Science Monitor: International Medical Journal of Experimental And Clinical Research, 2018, 24: 4379.
        Sample Type: Serum
      28. Publication: Ruan G, Xu J, Wang K, et al. Associations between knee structural measures, circulating inflammatory factors and MMP13 in patients with knee osteoarthritis[J]. Osteoarthritis and cartilage, 2018.
        Sample Type: Serum
      29. Publication: Li G Y , Liao Y , Hu J J , et al. Activation of NF-κB pathways mediating the inflammation and pulmonary diseases associated with atmospheric methylamine exposure[J]. Environmental Pollution, 2019, 252: 1216-1224.
        Sample Type: Cell supernatant
      30. Publication: Li G Y , Liao Y , Hu J J , et al. Activation of NF-κB pathways mediating the inflammation and pulmonary diseases associated with atmospheric methylamine exposure[J]. Environmental Pollution, 2019, 252: 1216-1224.
        Sample Type: Cell supernatant
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