Human IL-8 ELISA Kit (ab214030)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 1.8 pg/ml
- Range: 3.91 pg/ml - 250 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
-
Product name
Human IL-8 ELISA Kit
See all IL-8 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Overall 5 1.8% Inter-assay Sample n Mean SD CV% Overall 3 7.5% -
Sample type
Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
1.8 pg/ml -
Range
3.91 pg/ml - 250 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 99 93% - 103% Cell culture media 101 98% - 104% Hep Plasma 87 82% - 91% EDTA Plasma 88 84% - 92% Cit plasma 86 83% - 90% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
Human IL-8 ELISA Kit (ab214030) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IL-8 protein in cit plasma, edta plasma, hep plasma, serum, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IL-8 with 1.8 pg/ml sensitivity.
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. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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).
-
Notes
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.
-
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
-
Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human IL-8 Capture Antibody 1 x 600µl 10X Human IL-8 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 4BI 1 x 6ml Human IL-8 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
-
Function
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. -
Sequence similarities
Belongs to the intercrine alpha (chemokine CxC) family. -
Post-translational
modificationsSeveral N-terminal processed forms are produced by proteolytic cleavage after secretion from at least peripheral blood monocytes, leukcocytes and endothelial cells. In general, IL-8(1-77) is referred to as interleukin-8. IL-8(6-77) is the most promiment form. -
Cellular localization
Secreted. - Information by UniProt
-
Alternative names
- (Ala-IL-8)77
- (Ser-IL-8)72
- 9E3
see all -
Database links
- Entrez Gene: 3576 Human
- Omim: 146930 Human
- SwissProt: P10145 Human
- Unigene: 624 Human
Associated products
-
ELISA kits
-
KO cell lines
Images
-
Other - Human IL-8 ELISA Kit (ab214030)
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.
-
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.
-
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.
-
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.
-
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.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.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.
-
Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)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 (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.
-
Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)To learn more about the advantages of recombinant antibodies see here.
Datasheets and documents
-
SDS download
-
Datasheet download
References (3)
ab214030 has been referenced in 3 publications.
- Bardi GT et al. Detection of Inflammation-Related Melanoma Small Extracellular Vesicle (sEV) mRNA Content Using Primary Melanocyte sEVs as a Reference. Int J Mol Sci 20:N/A (2019). PubMed: 30870978
- Liu T et al. Self-seeding circulating tumor cells promote the proliferation and metastasis of human osteosarcoma by upregulating interleukin-8. Cell Death Dis 10:575 (2019). PubMed: 31366916
- Banach-Orlowska M et al. Cholesterol restricts lymphotoxin ß receptor-triggered NF-?B signaling. Cell Commun Signal 17:171 (2019). PubMed: 31878945