Human IL-8(Interleukin 8) ELISA Kit-Laboratory Biological Research Reagents Manufacturer for Life Science - Elabscience

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Human IL-8(Interleukin 8) ELISA Kit

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Price:   $495
Cat Number:  E-EL-H6008
Lead Time:   Order now, ship in 3 days

Reactivity:  Human

Detection Range:  7.81~500 pg/mL

Sensitivity:  4.69 pg/mL

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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-ELISA
Format 96T/48T
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-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
500 2.429
2.571
2.5 2.439
250 1.601
1.637
1.619 1.558
125 0.911
0.965
0.9379999999999999 0.877
62.5 0.46
0.488
0.474 0.413
31.25 0.272
0.273
0.273 0.212
15.63 0.173
0.188
0.181 0.12
7.81 0.119
0.124
0.122 0.061
0 0.063
0.058
0.061 --
sandwich-Ab-ELISA-Elabscience

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) 20.34 60.67 245.62 24.62 56.15 233.52
Standard deviation 1.21 2.80 12.26 1.39 2.80 9.69
C V (%) 5.95 4.62 4.99 5.65 4.99 4.15

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) 87-99 94
EDTA plasma (n=5) 95-102 98
Cell culture media (n=5) 98-106 102

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 (%) 97-110 93-105 102-113
Average (%) 102 101 105
1:4 Range (%) 93-107 92-101 99-111
Average (%) 99 95 105
1:8 Range (%) 87-97 90-103 93-103
Average (%) 92 95 97
1:16 Range (%) 93-104 94-110 91-102
Average (%) 97 102 94

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: Zhao Q, Liu J, Deng H, et al. Targeting Mitochondria-Located circRNA SCAR Alleviates NASH via Reducing mROS Output[J]. Cell, 2020.
    Sample Type: Cell culture medium
  2. Publication: Al‐Hakami A, Alqhatani S Q, Shaik S, et al. Cytokine physiognomies of MSCs from varied sources confirm the regenerative commitment post‐coculture with activated neutrophils[J]. Journal of Cellular Physiology, 2020.
    Sample Type: Cell culture supernatant
  3. 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
  4. 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 culture supernatant
  5. 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
  6. Publication: Bonvicini F, Pagnotta E, Punzo A, et al. Effect of Lactobacillus acidophilus Fermented Broths Enriched with Eruca sativa Seed Extracts on Intestinal Barrier and Inflammation in a Co-Culture System of an Enterohemorrhagic Escherichia coli and Human Intestinal Cells[J]. Nutrients, 2020, 12(10): 3064.
    Sample Type: Cell culture supernatant
  7. 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: Cell culture supernatant
  8. 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: Cell culture supernatant,Serum
  9. 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,Cell culture supernatant
  10. 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: Cell culture supernatant
  11. 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,Serum
  12. Publication: Xu J, Jia Z, Chen A, et al. Curcumin ameliorates Staphylococcus aureus-induced mastitis injury through attenuating TLR2-mediated NF-κB activation[J]. Microbial Pathogenesis, 2020, 142: 104054.
    Sample Type: Serum
  13. 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: Cell culture supernatant
  14. Publication: Huang W K, Xu Q Y, Zhou X, et al. Nec-1 attenuates inflammation and cytotoxicity induced by high glucose on THP-1 derived macrophages through RIP1[J]. Archives of Oral Biology, 2020, 118: 104858.
    Sample Type: Cell culture supernatant
  15. 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
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