Human IL-8 ELISA

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ELH-IL8-1

RayBio® Human IL-8 (CXCL8) ELISA Kit for cell culture supernatants, plasma, and serum samples.

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Antigen Information

Accession Number:
  • P10145
Gene ID:
  • 3576
Gene Symbols:
  • CXCL8
  • IL8
Protein Name & Synonyms:
Interleukin-8 (IL-8) (C-X-C motif chemokine 8) (Chemokine (C-X-C motif) ligand 8) (Emoctakin) (Granulocyte chemotactic protein 1) (GCP-1) (Monocyte-derived neutrophil chemotactic factor) (MDNCF) (Monocyte-derived neutrophil-activating peptide) (MONAP) (Neutrophil-activating protein 1) (NAP-1) (Protein 3-10C) (T-cell chemotactic factor) [Cleaved into: MDNCF-a (GCP/IL-8 protein IV) (IL8/NAP1 form I) Interleukin-8 ((Ala-IL-8)77) (GCP/IL-8 protein II) (IL-8(1-77)) (IL8/NAP1 form II) (MDNCF-b) IL-8(5-77) IL-8(6-77) ((Ser-IL-8)72) (GCP/IL-8 protein I) (IL8/NAP1 form III) (Lymphocyte-derived neutrophil-activating factor) (LYNAP) (MDNCF-c) (Neutrophil-activating factor) (NAF) IL-8(7-77) (GCP/IL-8 protein V) (IL8/NAP1 form IV) IL-8(8-77) (GCP/IL-8 protein VI) (IL8/NAP1 form V) IL-8(9-77) (GCP/IL-8 protein III) (IL8/NAP1 form VI)]
Target Species:
  • Human

Assay Format

Detection Range:
1 pg/ml - 600 pg/ml
Recommended Dilution (serum & plasma):
2 fold
Sensitivity:
1 pg/ml

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Specificity:
This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, 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 (OB), 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.
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes
  • 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

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • 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
Other Materials Required
  • 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
Protocol Outline
  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.

Storage/Stability

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.
  1. Dominguez C., McCampbell K., David J., Palena C. Neutralization of IL-8 decreases tumor PMN-MDSCs and reduces mesenchymalization of claudin-low triple-negative breast cancer. JCI Insight. 2017 Nov 2;2(21). pii: 94296. doi: 10.1172/jci.insight.94296.
    Species: Human
    Sample type: Conditioned Media
  2. Xu D., Li J., Huang X., et al. Impact of IL-8-251A/T gene polymorphism on severity of disease caused by enterovirus 71 infection. Arch Virol. 2016 Jan;161(1):203-7. doi: 10.1007/s00705-015-2645-2.
    Species: Human
    Sample type: Plasma
  3. Duckworth C., Zhang L., Carroll S., et al. Overexpression of GAB2 in ovarian cancer cells promotes tumor growth and angiogenesis by upregulating chemokine expression. Oncogene. 2016 Aug 4;35(31):4036-47. doi: 10.1038/onc.2015.472.
    Species: Human
    Sample type: Conditioned Media
  4. Kuntz S., Asseburg H., Dold S., et al. Inhibition of low-grade inflammation by anthocyanins from grape extract in an in vitro epithelial-endothelial co-culture model. Food Funct. 2015 Apr;6(4):1136-49. doi: 10.1039/c4fo00755g.
    Species: Human
    Sample type: Conditioned Media
  5. Perrigue P., Silva M., Warden C., et al. The histone demethylase jumonji coordinates cellular senescence including secretion of neural stem cell-attracting cytokines. Mol Cancer Res. 2015 Apr;13(4):636-50. doi: 10.1158/1541-7786.MCR-13-0268
    Species: Human
    Sample type: Conditioned Media
  6. Moore Q., Smith C., Wolf S., et al. Examining the immune response of pneumolysin exposed human corneal epithelial cells in a keratitis model. BIOS 85(4):199-206. 2014. doi: http://dx.doi.org/10.1893/0005-3155-85.4.199
    Species: Human
    Sample type: Conditioned Media
  7. Li J., Lin A., Yu C., et al. Association of Enterovirus 71 encephalitis with the interleukin-8 gene region in Chinese children. Infectious Diseases Volume 47, Issue 6, 2015. DOI:10.3109/00365548.2015.1007473
    Species: Human
    Sample type: Plasma
  8. Gao S., Guan S., Li H., et al. Ameliorating effects of low tidal volume ventilation with associated hypercapnia on pneumoperitoneum-induced lung injury by inhibition of Toll-like receptor 4. Int J Clin Exp Med. 2015 Feb 15;8(2):1814-23.
    Species: Rat
    Sample type: BAL Fluid
  9. Zhou X., Gu J., Gu Y., et al. Human Umbilical Cord-Derived Mesenchymal Stem Cells Improve Learning and Memory Function in Hypoxic-Ischemic Brain-Damaged Rats via an IL-8-Mediated Secretion Mechanism Rather than Differentiation Pattern Induction. Cell Physiol Biochem. 2015;35(6):2383-401. doi: 10.1159/000374040
    Species: Human
    Sample type: Conditioned Media
  10. Ikeno H., Apel M., Zouboulis C., et al. l-Ascorbyl-2-phosphate attenuates NF-?B signaling in SZ95 sebocytes without affecting IL-6 and IL-8 secretion. Arch Dermatol Res. 2015; 307(7): 595-605. doi: 10.1007/s00403-015-1565-z
    Species: Human
    Sample type: Conditioned Media
Easy to follow and works exactly as expected.

Biocogent, LLC
Best Elisa. Easy to use instructions very helpful for new users.

Wvu
It is very user-friendly well written protocol. Easy to use and get results in one day.

University of Illinois at Chicago
I used this kit to measure IL-8 in culture supernatants after HIV treatment. The results look good and there was no high background in my negative controls.
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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.