Human IL-8 ELISA Kit (A2982)

Name: Human IL-8 ELISA Kit
Brand: Antibodies.com
SKU: A2982
Price: 300.0 GBP
Availability: InStock

1 Citation

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Name

Human IL-8 ELISA Kit

Description

Human IL-8 ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human IL-8 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.

Assay Type

Sandwich (quantitative)

Principle of Assay

Human IL-8 ELISA Kit (A2982) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human IL-8 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. An antibody specific for IL-8 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the IL-8 present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-IL-8 Antibody, which binds the captured IL-8 present in each well. HRP-Avidin conjugate is then added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of IL-8 captured in each well. The concentration of IL-8 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.

Platform

Pre-coated Microplate (12 x 8 well strips)

Detection Type

Colourmetric

Instrument

Colorimetric Microplate Reader

Sample Type

Serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.

Sensitivity

3.6 pg/ml

Product Range

7.81-500 pg/ml

Assay Time

4h 30m

Reactivity

Human

Recovery

Sample Type	n	Range	Average
Serum	5	80% - 102%	91%
EDTA Plasma	5	81% - 100%	90%
Heparin Plasma	5	80% - 89%	84%

Linearity

Sample Type	1:2	1:4	1:8	1:16
Serum (n=5)	87-91%	87-107%	74-101%	92-97%
EDTA Plasma (n=5)	90-105%	84-101%	90-101%	79-108%
Heparin Plasma (n=5)	84-95%	92-105%	82-105%	89-91%

Precision

Intra-assay: CV < 10%, Inter-assay: CV < 12%

General Notes

Information online is representative. Always refer to the Product Manual inside the kit.

Storage

Store kit components at +4°C or -20°C as instructed. Do not use past expiration date!

Synonyms

C-X-C motif chemokine 8, C-X-C motif) ligand 8, Chemokine, CXCL8, Emoctakin, GCP-1, Granulocyte chemotactic protein 1, IL8, Interleukin-8, MDNCF, Monocyte-derived neutrophil chemotactic factor

Product Note

Information online is representative. Always refer to the Product Manual inside the kit.

Disclaimer

This product is for research use only. It is not intended for diagnostic or therapeutic use.

Kit Components

Item	Quantity	Storage
Pre-Coated 96 Well Microplate	12 x 8 Well Strips	-20°C
Lyopholized Standard	2 Vials	-20°C
Detection Solution A	120μl	-20°C
Detection Solution B	120µl	-20°C
Wash Buffer (30X)	20ml	+4°C
Sample Dilution Buffer	45ml	-20°C
TMB Substrate	9ml	+4°C
Stop Solution	6ml	+4°C
Plate Sealers	5 Adhesive Strips	-

Required Materials

The following materials are not included in the kit, but are required to run this assay:

Microplate reader capable of measuring absorbance at 450nm ± 10nm.
Squirt bottle, multichannel pipette reservoir, or automated microplate washer.
Graph paper or computer software capable of generating logarithmic functions.
Test tubes and Eppendorf tubes to prepare standards and samples.
Multi- and single-channel pipettes and sterile pipette tips.
Deionized or distilled water.
Absorbent paper towels.
37°C incubator.

Important Notes

Sample concentrations should be estimated before being used in the assay and a proper dilution factor should be selected to make the diluted target protein concentration fall in the optimal detection range of this kit.
Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.
Samples should be brought to room temperature before starting the assay.
This ELISA kit is intended for the analysis of biological samples and has not been validated for purified/recombinant proteins, industrial, or commercial materials. Performance with such samples cannot be guaranteed, as differences in expression, sequence, tags, folding, post-translational modifications, purity, activity, or buffer composition may affect assay results. For further guidance, please contact our technical support team.

Bring all reagents and samples to room temperature before use.

We recommend that all standards and samples be assayed in duplicate.

Step 1

Prepare all reagents, samples, and working standards as directed in the previous sections.

Step 2

Determine the number of microplate strips required for the assay and insert them into the plate frame. Unused strips should be stored in a foil pouch at -20°C.

Step 3

Add 100μl of standard solutions to the standard wells. Note: A plate layout is provided to record standards and samples assayed.

Step 4

Add 100μl of samples to the sample wells.

Step 5

Cover with an adhesive plate sealer provided and incubate at 37°C for 2 hours.

Step 6

Remove the plate sealer and aspirate the liquid from the plate.

Step 7

Add 100μl of working Detection Solution A to each well.

Step 8

Cover with a new adhesive plate sealer and incubate at 37°C for 60 minutes.

Step 9

Remove the plate sealer, aspirate the liquid from the plate, and wash the plate three times with Wash Buffer. Soak wells with 300μl of Wash Buffer for 1-2 minutes each time. Note: Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Blot the plate onto paper towels or other absorbent material. Do not let the wells dry completely at any time!

Step 10

Add 100μl of working Detection Solution B to each well.

Step 11

Cover with a new adhesive plate sealer and incubate at 37°C for 60 minutes.

Step 12

Remove the plate sealer, aspirate the liquid from the plate, and wash the plate five times with Wash Buffer. Soak wells with 300μl of Wash Buffer for 1-2 minutes each time.

Step 13

Add 90μl of TMB Substrate to each well. Cover with a new plate sealer and incubate at 37°C in the dark for 15-25 minutes (do not exceed 30 minutes). Note: This incubation time is for reference only, the optimal reaction time should be determined by the end-user and can be shortened or extended according to the actual color change. The reaction may be terminated when a gradient is apparent in the standard wells.

Step 14

Add 50μl of Stop Solution to each well. The color in the wells should change from blue to yellow immediately. Note: If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.

Step 15

Determine the optical density (O.D. value) of each well immediately using a microplate reader set to 450 nm.

Scientific Validation DataValidation Data (1)

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Standard Curve - Human IL-8 ELISA Kit (A2982)

Representative standard curve for Human IL-8 ELISA Kit (A2982).

Product Citations (1)

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Anisakis simplex larvae viability and potential pathogenicity in vitro is controlled by the essential oil from sea fennel (Crithmum maritinum L., Apiaceae)

This publication cites Human IL-8 ELISA Kit (A2982).

Article Abstract

Essential oils, natural volatile products extracted from aromatic plants, exhibit a wide range of biological activities, including antimicrobial and antiparasitic properties. Anisakis simplex is a parasitic nematode commonly found in marine fish, which poses a significant health risk to humans through the consumption of raw or undercooked seafood, leading to anisakiasis, a zoonotic disease with both gastrointestinal and allergic manifestations. In this study, the impact of Crithmum maritimum L. (Apiaceae) essential oil on this nematode was evaluated in vitro. The chemical composition of C. maritimum essential oil was determined by gas chromatography-mass spectrometry (GC-MS). The analysis revealed ?-terpinene, limonene, and sabinene as the major components. The nematicidal activity, larval penetration ability, resistance to acidic pH, and inhibition of acetylcholinesterase were evaluated. The essential oil effects on interleukin production and cytotoxicity in HepG2 and Caco-2 cells were also assessed. C. maritimun exhibited larvicidal activity, with LD50 values of 619.11 µg mL^-1 at 24 h and 353.95 µg mL^-1 at 48 h. Additionally, it showed no significant cytotoxicity in HepG2 and Caco-2 cell lines at the concentrations tested. In Caco-2 cells stimulated with crude Anisakis extract, the essential oil modulated interleukin production. Furthermore, it demonstrated in vitro inhibitory activity against 5-lipoxygenase and phospholipase A2, with IC50 values of 99.35 and 253.31 µg mL^-1, respectively. The traditional culinary use of C. maritimum, together with the observed anti-Anisakis activity in our study, suggest this essential oil may offer a promising complementary approach within broader integrated food safety strategies, especially in Anisakis control.