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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


Images

Sandwich ELISA - Human IL-8 ELISA Kit (AB214030), expandable thumbnail
  • Sandwich ELISA - Human IL-8 ELISA Kit (AB214030), expandable thumbnail
  • Sandwich ELISA - Human IL-8 ELISA Kit (AB214030), expandable thumbnail
  • Sandwich ELISA - Human IL-8 ELISA Kit (AB214030), expandable thumbnail
  • Sandwich ELISA - Human IL-8 ELISA Kit (AB214030), expandable thumbnail

Publications

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

3.91 - 250 pg/mL

Assay time

1h 30m

Sensitivity

= 1.8 pg/mL

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Reactivity data

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

What's included?

1 x 384 Tests
Components
10X Human IL-8 Capture Antibody
1 x 600 µL
10X Human IL-8 Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
384 well CaptSure™ microplates
1 x 1 Unit
Antibody Diluent 4BI
1 x 6 mL
Human IL-8 Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
2 x 250 mL
Stop Solution
2 x 12 mL
TMB Development Solution
2 x 12 mL

Recommended products

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

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

3.91 - 250 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 1.8 pg/mL

Precision

Intra assay

Sample

Overall

n
5
mean
-
SD
-
C.V.

1.8

Inter assay

Sample

Overall

n
3
mean
-
SD
-
C.V.

7.5

Recovery

Sample specific recovery

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 %

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

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.

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-1.

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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10 product images

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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.

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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.

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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 (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    Example of human IL-8 standard curve.

    Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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 (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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.

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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.

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - Human IL-8 ELISA Kit (ab214030)

    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), expandable thumbnail

    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 (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.

  • Sandwich ELISA - Human IL-8 ELISA Kit (ab214030), expandable thumbnail

    Sandwich ELISA - 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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