Human IL-8(Interleukin 8) ELISA Kit

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    • 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-H0048

      Synonyms:IL8, CXCL8, GCP-1, GCP1, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP-1, NAP1, 3-10C, AMCF-I, K60

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

      Reactivity: Human

      Detection Range: 31.25~2000 pg/mL

      Sensitivity: 18.75 pg/mL

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

      Add to cart Compare Bulk request Manual

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human IL-8 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-8. 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-8 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-8, 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-8. You can calculate the concentration of Human IL-8 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 31.25—2000 pg/mL
      Sensitivity 18.75 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human IL-8 in samples. No significant cross-reactivity or interference between Human IL-8 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
      2000 2.477
      2.491
      2.484 2.397
      1000 1.588
      1.634
      1.611 1.524
      500 0.979
      0.943
      0.961 0.874
      250 0.47
      0.482
      0.476 0.389
      125 0.264
      0.264
      0.264 0.177
      62.5 0.201
      0.177
      0.189 0.102
      31.25 0.135
      0.143
      0.139 0.052
      0 0.086
      0.088
      0.087 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL-8 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-8 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) 111.10 220.20 878.40 108.90 237.70 865.90
      Standard deviation 7.10 12.80 40.40 7.40 12.40 46.80
      C V (%) 6.39 5.81 4.60 6.80 5.22 5.40

      Recovery

      The recovery of Human IL-8 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) 89-106 97
      EDTA plasma (n=5) 94-106 101
      Cell culture media (n=5) 90-104 97

      Linearity

      Samples were spiked with high concentrations of Human IL-8 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 (%) 91-104 96-109 92-106
      Average (%) 97 102 98
      1:4 Range (%) 100-111 81-93 98-114
      Average (%) 106 87 104
      1:8 Range (%) 96-107 84-95 93-107
      Average (%) 102 90 98
      1:16 Range (%) 95-110 83-93 91-105
      Average (%) 103 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

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

      2.Remove the liquid.

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

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

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

      6.Add 50 μL Stop Solution.

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

      Citations

      1. 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
      2. Publication: Tang Y, Jin X, Xiang Y, et al. The lncRNA MALAT1 Protects The Endothelium Against ox-LDL-Induced Dysfunction Via Upregulating The Expression Of The miR-22-3p Target Genes CXCR2 And AKT[J]. Febs Letters, 2015, 589(20): 3189-3196.
        Sample Type: culture supernatant
      3. Publication: Yan Z, Wang W, Wu Y, et al. Zinc Oxide Nanoparticle-Induced Atherosclerotic Alterations In Vitro And In Vivo[J]. International Journal of Nanomedicine, 2017, 12: 4433-4442.
        Sample Type: culture supernatant and Serum
      4. 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
      5. 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
      6. 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
      7. 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: Human
      8. 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: Human
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