Human IL-8 ELISA by RayBiotech Inc.

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

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; Cell Lysates; Plasma; Serum; Tissue Lysates

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

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.