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Product Specifications

Analytical sensitivity

<0.01 pg/mL

Assay range

0.0128 - 5000 pg/mL

Sample type/volume

Serum
2-5 µL
Plasma
2-5 µL
Supernatant
2-5 µL

Hands-on time

30 min

Time-to-result

2 hr

Homogenous (no wash)

Yes

Instrument

qPCR Instruments

Product size

96 Tests

Contents

Assay Dilution Buffer
Antibody-Conjugate A
Antibody-Conjugate B
Antibody-Conjugate Dilution Buffer
Standard
Master Mix
Ligase

Shipping conditions

Dry ice

Storage

-20°C

Protein name

IL-8

Species (tested)

Human

Assay kit format

Proximity-based Immunoassay

Label or dye

FAM™

About This Kit

The ProQuantum Human IL-8 Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human IL-8 protein in small sample volumes. Utilizing proximity ligation assay (PLA) technology, the assay combines the analyte specificity of high-affinity antibody-antigen binding with the signal detection and amplification capabilities of real-time PCR to achieve a simple yet powerful next-generation protein quantitation platform. A user-friendly workflow combined with intuitive Cloud-based software for analytics enables sample-to-answer in just 2 hours.

• High sensitivity-detect low levels of protein with greater sensitivity than traditional methods

• Broad dynamic range-≥5 logarithmic units, minimizing sample dilutions to ensure they fall within the range

• Small sample consumption-use 2-5 µL of sample (compared to 75 µL for triplicate wells with other methods)

• Fast, easy workflow-2 hours from sample to answer, with no wash steps

• No proprietary instrument to purchase-runs on any real-time PCR instrument

ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody-analyte binding, the two DNA oligos are brought into close proximity, which allows for ligation of the two strands and subsequent creation of a template strand for amplification. This platform leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan real-time PCR technology (Figure 1).

The assay workflow is fast and easy-2 steps in 2 hours. There are a total of 7 components in each kit (Figure 2). First, mix the antibody-conjugates, dilute the sample, and create the standard curve in a working plate. Then, using a multi-channel pipette, add the antibody-conjugates and sample (or standard) into the wells of a PCR plate and incubate for 1 hour. Combine the master mix and ligase and add to the wells of the PCR plate, then run the plate on any qPCR instrument. After the run is complete, import the results file into the ProQuantum cloud-based software at https://apps.thermofisher.com/apps/proquantum. Using this software, the data can be analyzed easily to obtain protein concentration values. The software allows you to set up standard curves, design plate layouts, set up customized assay instructions, and obtain robust statistical group-wise comparisons.

Target information

Interleukin 8 (IL-8, CXCL8) is a 72 amino acid pro-inflammatory factor which belongs to the CXC subfamily of chemokines, and are bound by the cell surface receptors IL-8RA and IL-8RB. IL-8 functions as a chemoattractant and potent angiogenic factor. The expression and secretion of IL-8 can be induced by diverse inflammatory stimuli in many cells, including macrophages and endothelial cells. In endothelial cells, IL-8 is present in storage vesicles called Weibel-Palade bodies. IL-8, first isolated from osteosarcoma cells, contains the ELR-motif (N-terminal Glu-Leu-Arg amino acid sequence) and signals through the CXCR1 and CXCR2 receptors. Previous nomenclature for IL-8 includes neutrophil activating protein 1 (NAP-1), granulocyte chemotactic protein 1 (GCP-1), monocyte-derived neutrophil-activating peptide (MONAP) and protein 3-10C. IL-8 and ten other members of the CXC chemokine gene family form a chemokine gene cluster in a region mapped to chromosome 4q. Cancer studies have demonstrated a role for IL-8 in the angiogenesis and growth of tumours, and IL-8 is believed to play a role in the pathogenesis of bronchiolitis, a common respiratory tract disease caused by viral infection.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Gene aliases : CXCL8, GCP-1, GCP1, IL8, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP-1, NAP1

Gene ID : (Human) 3576

Gene symbol : CXCL8

Protein Aliases : alveolar macrophage chemotactic factor I, beta endothelial cell-derived neutrophil activating peptide, beta-thromboglobulin-like protein, C-X-C motif chemokine 8, Chemokine (C-X-C motif) ligand 8, Emoctakin, GCP-1, Granulocyte chemotactic protein 1, IL-8, interleukin 8, Interleukin-8, lung giant cell carcinoma-derived chemotactic protein, lymphocyte derived neutrophil activating peptide, MDNCF, MONAP, Monocyte-derived neutrophil chemotactic factor, Monocyte-derived neutrophil-activating peptide, NAP-1, neutrophil-activating peptide 1, Neutrophil-activating protein 1, Protein 3-10C, small inducible cytokine subfamily B, member 8, T-cell chemotactic factor, tumor necrosis factor-induced gene 1

UniProt ID (Human) P10145

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