Human IL-8 ELISA Kit (ab214030)
- Datasheet
- References (2)
- Protocols
Overview
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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 IL8 SimpleStep ELISA kit (ab174442) has been re-developed with new capture and detector antibodies. This new kit will have the same name but a different product number (ab214030). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA® platform that provide a higher sensitivity when quantification of IL8 in human serum, plasma and cell culture supernatants.
IL-8 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-8 protein in serum, plasma and cell culture supernatant.
The 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. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
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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.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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
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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
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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
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ELISA kits
Images
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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.
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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.
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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.
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Example of human IL-8 standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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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.
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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).
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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.
Protocols
References
This product has been referenced in:
- 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). Read more (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). Read more (PubMed: 31366916) »
