Human IL-8 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human IL-8 with a sensitivity of 0.45 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
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- International journal of biological sciences 18:3337-33572022Neutrophil extracellular traps mediate mA modification and regulates sepsis-associated acute lung injury by activating ferroptosis in alveolar epithelial cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHao Zhang,Jinlong Liu,Yilu Zhou,Mengdi Qu,Yanghanzhao Wang,Kefang Guo,Ruling Shen,Zhirong Sun,Juan P Cata,Shuofei Yang,Wankun Chen,Changhong MiaoPubMed 35637949
- Communications biology 5:3792022Silencing of STE20-type kinase STK25 in human aortic endothelial and smooth muscle cells is atheroprotective.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEmmelie Cansby,Sima Kumari,Mara Caputo,Ying Xia,Rando Porosk,Jonathan Robinson,Hao Wang,Britt-Marie Olsson,Josefine Vallin,Julie Grantham,Ursel Soomets,L Thomas Svensson,Carina Sihlbom,Hanns-Ulrich Marschall,Andreas Edsfeldt,Isabel Goncalves,Margit MahlapuuPubMed 35440683
- The Journal of investigative dermatology 142:1670-1681.e122021Nicotinamide Prevents UVB- and Oxidative Stress‒Induced Photoaging in Human Primary Keratinocytes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChristina Yan Ru Tan,Chye Ling Tan,Toby Chin,Malgorzata Morenc,Chin Yee Ho,Holly A Rovito,Ling Shih Quek,Ai Ling Soon,John S Y Lim,Oliver Dreesen,John E Oblong,Sophie BellangerPubMed 34740582
Key facts
- Detection method
- Fluorescent
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.49 - 2000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 0.45 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
Chemotactic factor that mediates inflammatory response by attracting neutrophils, basophils, and T-cells to clear pathogens and protect the host from infection (PubMed:18692776, PubMed:7636208). Also plays an important role in neutrophil activation (PubMed:2145175, PubMed:9623510). Released in response to an inflammatory stimulus, exerts its effect by binding to the G-protein-coupled receptors CXCR1 and CXCR2, primarily found in neutrophils, monocytes and endothelial cells (PubMed:1840701, PubMed:1891716). G-protein heterotrimer (alpha, beta, gamma subunits) constitutively binds to CXCR1/CXCR2 receptor and activation by IL8 leads to beta and gamma subunits release from Galpha (GNAI2 in neutrophils) and activation of several downstream signaling pathways including PI3K and MAPK pathways (PubMed:11971003, PubMed:8662698).
Alternative names
IL8, CXCL8, Interleukin-8, IL-8, C-X-C motif chemokine 8, Chemokine (C-X-C motif) ligand 8, Emoctakin, Granulocyte chemotactic protein 1, Monocyte-derived neutrophil chemotactic factor, Monocyte-derived neutrophil-activating peptide, Neutrophil-activating protein 1, Protein 3-10C, T-cell chemotactic factor, GCP-1, MDNCF, MONAP, NAP-1
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Human IL-8 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human IL-8 with a sensitivity of 0.45 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.49 - 2000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 0.45 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Supernatant | n 5 | C.V. 1.8 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Supernatant | n 3 | C.V. 7.5 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Serum | Average % = 99 | Range 93 - 103 % |
Sample type EDTA Plasma | Average % = 88 | Range 84 - 92 % |
Sample type Heparin Plasma | Average % = 87 | Range 82 - 91 % |
Sample type Citrate plasma | Average % = 86 | Range 83 - 90 % |
Sample type Cell culture media | Average % = 101 | Range 98 - 104 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
IL-8 in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-8 protein in human serum, plasma, and cell culture supernatant.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.
The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
IL-8 is a chemotactic factor that attracts neutrophils, basophils, and T cells, but not monocytes. It is also involved in neutrophil activation. It is released from several cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL-8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77), respectively
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
IL-8 also known as CXCL8 is a cytokine with a molecular weight of approximately 8 kDa. It belongs to the CXC chemokine family and plays a role in the recruitment of neutrophils to sites of inflammation. Cells like monocytes macrophages and endothelial cells express IL-8 after activation by pro-inflammatory signals. The protein exhibits high expression in cells associated with the immune response and inflammation serving as a signaling molecule between these cell types.
- Biological function summary
IL-8 functions as a chemoattractant for neutrophils and lymphocytes facilitating their movement towards the site of infection or injury. It does not form part of a larger protein complex but operates individually to enhance immune cell migration. IL-8 possesses unique binding motifs allowing it to interact with specific receptors namely CXCR1 and CXCR2 on target cells. This binding triggers cellular responses leading to effective immune surveillance and response to inflammatory stimuli.
- Pathways
IL-8 operates within important inflammatory and immune response pathways. It forms a part of the NF-κB signaling cascade which activates in response to stress signals promoting the expression of other inflammatory mediators. Additionally it engages in the mitogen-activated protein kinase (MAPK) pathway influencing cellular responses such as proliferation and differentiation. The interaction of IL-8 with these pathways highlights its role in modulating immune responses and highlights its interaction with other proteins such as TNF-α and IL-1β.
- Associated diseases and disorders
IL-8 shows a strong association with conditions like rheumatoid arthritis and chronic obstructive pulmonary disease (COPD). It acts as a biomarker for these diseases often correlating with disease severity and inflammation levels. In rheumatoid arthritis IL-8 contributes to joint inflammation and degradation while in COPD it enhances airway inflammation and tissue damage. The cytokine's involvement in these diseases connects it with other inflammatory mediators such as MMPs and leukotrienes which collectively exacerbate disease pathology.
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5 product images
Sandwich ELISA - Human IL-8 ELISA Kit, Fluorescent (ab229402)
Example of human IL-8 standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human IL-8 ELISA Kit, Fluorescent (ab229402)
Interpolated concentrations of native IL-8 in human serum.
The concentrations of IL-8 were measured in duplicates, interpolated from the IL-8 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-8 concentration was determined to be 63.2 pg/mL in serum.
Sandwich ELISA - Human IL-8 ELISA Kit, Fluorescent (ab229402)
Interpolated concentrations of spiked IL-8 in human plasma samples.
The concentrations of IL-8 were measured in duplicates, interpolated from the IL-8 standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (EDTA) 50%, plasma (citrate) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
Sandwich ELISA - Human IL-8 ELISA Kit, Fluorescent (ab229402)
Serum from ten individual healthy human female donors was measured in duplicate.
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-8 concentration was determined to be 61.6 pg/mL with a range of 62 – 174.9 pg/mL; two individuals (Donor #3 and Donor #9) measured below the minimal detectable dose.
Sandwich ELISA - Human IL-8 ELISA Kit, Fluorescent (ab229402)
Interpolated concentrations of native IL-8 in human PBMC cell culture supernatant.
The concentrations of IL-8 were measured in duplicates, interpolated from the IL-8 standard curves and corrected for sample dilution. Undiluted samples are as follows: unstimulated 1:400 and stimulated 1:400. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Measured values were interpolated from the IL-8 Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. The mean IL-8 concentration was determined to be 2.2 ng/mL in unstimulated, 119.7ng/mL in stimulated, and undetectable in media.
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