The human IL-8 ELISA kit is a single-wash, 90-minute SimpleStep ELISA® assay designed for the quantification of human interleukin-8 (IL-8) with a sensitivity of 1.8 pg/mL. The assay employs a simple Mix-Wash-Read protocol with just one incubation and wash step.Easy-to-use, rapid, sensitive human interleukin-8 (IL-8) sandwich ELISA kit:- Colorimetric assay with a 450 nm readout; compatible with any standard plate reader- Available in different formats to meet various throughput needs, including 384-well plates for higher throughput- Broad sample compatibility – suitable for serum, plasma (heparin, citrate, EDTA), cell culture supernatants, and cerebrospinal fluid- Cited in over 30 publications
Colorimetric
Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
Sandwich (quantitative)
Human
3.91 - 250 pg/mL
1h 30m
= 1.8 pg/mL
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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).
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
The human IL-8 ELISA kit is a single-wash, 90-minute SimpleStep ELISA® assay designed for the quantification of human interleukin-8 (IL-8) with a sensitivity of 1.8 pg/mL. The assay employs a simple Mix-Wash-Read protocol with just one incubation and wash step.Easy-to-use, rapid, sensitive human interleukin-8 (IL-8) sandwich ELISA kit:- Colorimetric assay with a 450 nm readout; compatible with any standard plate reader- Available in different formats to meet various throughput needs, including 384-well plates for higher throughput- Broad sample compatibility – suitable for serum, plasma (heparin, citrate, EDTA), cell culture supernatants, and cerebrospinal fluid- Cited in over 30 publications
Colorimetric
Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
Sandwich (quantitative)
Human
3.91 - 250 pg/mL
1h 30m
Pre-coated microplate (12 x 8 well strips)
= 1.8 pg/mL
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Overall | n 5 | mean - | SD - | C.V. 1.8 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Overall | n 3 | mean - | SD - | C.V. 7.5 |
Sample type | Average % | Range |
---|---|---|
Sample type Cell culture media | Average % = 101 | Range 98 - 104 % |
Sample type Cerebral Spinal Fluid | Average % = 89 | Range 84 - 94 % |
Sample type Citrate plasma | Average % = 86 | Range 83 - 90 % |
Sample type EDTA Plasma | Average % = 88 | Range 84 - 92 % |
Sample type Heparin Plasma | Average % = 87 | Range 82 - 91 % |
Sample type Serum | Average % = 99 | Range 93 - 103 % |
Blue Ice
+4°C
+4°C
+4°C
Human IL-8 ELISA Kit ab214030 is a rapid single-wash 90-min ELISA kit to measure Human IL-8 in cerebral spinal fluid (CSF), cell culture supernatant, serum and plasma. This SimpleStep ELISA® sensitivity is 1.8 pg/mL.
How the assay works
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.
Assay protocol summary
Human IL-8 ELISA Kit ab214030 protocol summary:
1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hour at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm
Alternative assay kits
Need to design your own immunoassay? We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:
- Human/Monkey IL-8 Antibody Pair - BSA and Azide free Human/Monkey IL-8 Antibody Pair - BSA and Azide free ab241839,
- Anti-IL-8 antibody [EPR19358-37] - BSA and Azide free (Capture) Anti-IL-8 antibody [EPR19358-37] - BSA and Azide free (Capture) ab242515,
- Anti-IL-8 antibody [EPR19358-108] - BSA and Azide free (Detector) Anti-IL-8 antibody [EPR19358-108] - BSA and Azide free (Detector) ab242983
Getting the best performance
ASSAY SPECIFICITY
This kit recognizes both native and recombinant human IL-8 protein in serum, plasma, and cell culture supernatant samples only.
Cell and tissue extract samples have not been tested with this kit.
SPECIES REACTIVITY
This kit recognizes human IL-8 protein.
Other species reactivity was determined by measuring 100% mouse, rat, and bovine serum samples, interpolating the IL-8 protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the IL-8 protein concentration in human serum assayed at the same dilution.
Reactivity < 3% was determined for the following species: Mouse, Rat, Cow
CALIBRATION
This immunoassay is calibrated against a highly purified human IL-8. The NIBSC/WHO unclassified purified human IL-8 preparation 89/520 was evaluated in this kit.
The dose response curve of the unclassified standard IL-8 parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep Human IL-8 kit to approximate NIBSC IU/mL units, use the equation below.
NIBSC (89/250) approximate value (IU/mL) = 0.0042 x SimpleStep Human IL-8 value (pg/mL).
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.
Save your precious samples
Save your precious samples: our 384-well Human IL-8 ELISA Kit only requires a maxmum of 12.5µL
How other researchers are using this kit
Human IL-8 ELISA Kit ab214030 has been used to assess inflammation in different physio-pathological contexts such as traumatic brain injury (1), pulmonary disease (2), and osteoarthritis (3).
References: 1-Jindan Gao et al. 2022, PMID: 36247290, 2-Rui Zheng et al. 2022, PMID: 35191419, 3-Zhi Lv et al. 2021, PMID: 34082824
Related and recommended products
Human IL-8 ELISA Kit ab214030 is commonly used to measure IL-8 as a marker of inflammation.
Other ELISA kits to measure inflammatory markers include:
- Human TNF alpha ELISA kit Human TNF alpha ELISA Kit ab181421
- Human IL-6 ELISA kit Human IL-6 ELISA Kit ab178013
- Human IL-1 beta ELISA kit Human IL-1 beta ELISA Kit ab214025
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.
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.
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-1.
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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Human IL-8 standard curve comparison data.
Standard curve comparison between human IL-8 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA® kit shows a 3-fold increase in sensitivity.
Example of human IL-8 standard curve in Sample Diluent NS.
The IL-8 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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.
Example of human IL-8 standard curve.
Background-subtracted data values (mean +/- SD) are graphed.
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.
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. Mean of duplicate values +/-SD are graphed. The mean IL-8 concentration was determined to be 2.2 ng/mL in unstimulated, 119.7ng/mL in stimulated, and undetectable in media.
Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Native IL-8 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Recombinant IL-8 was spiked into the following biological samples and diluted in a 2-fold dilution series in Sample Diluent NS.
Human IL-8 concentration was interpolated from the standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-8 ELISA kit (ab214030). Wild-type PC-3 cells and CXCL8 knockout PC-3 cells (Human CXCL8 knockout PC-3 cell line ab273743) were assessed in duplicate (n=2) and were either treated with 2 μg/ml LPS for 6 hours to induce expression of IL-8 or not treated with LPS. Data are represented as the mean and error bars represent standard deviation.
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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