Sample Type: serum, plasma, Tissue homogenate and Other biological samples
Product Status: Available
NEKA-EH4342 Human IL-8 (Interleukin 8) ELISA Kit
Principle of Human IL 8 Interleukin 8 ELISA Kit Test
The Human IL-8 Interleukin 8 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in-vitro enzyme-linked immunosorbent assay for the quantitative measurement of human IL-8 in serum, plasma, and cell culture supernatants. This assay employs the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to IL-8. Standards or samples are added to the wells,alongside a biotin-conjugated antibody specific for IL-8. Then, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. After TMB substrate solution is added, only wells that contain IL-8, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated, and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.
Importance of the Test
IL-8, also known as Interleukin 8, is a chemokine produced by macrophages and other cell types such as epithelial cells, airway smooth muscle cells, and endothelial cells. Il-8, with its sister molecule IL-6, are the first cytokines to be released by macrophages when they encounter an infection. Elevated levels of IL-8 are often associated with inflammation and has been noted in several disease conditions like colitis, periodontitis, bronchiectasis etc. This assay provides a reliable and accurate quantitative measurement of IL-8 levels which can be critical in clinical diagnostic for certain conditions.
List of Advantages
No significant cross-reactivity or interference between human IL-8 and analogues.
Broad detection range and high sensitivity.
Instant ELISA kit reduces experiment time.
Stable antibodies. The loss of detection capability is less than 5% within the expiration period.
For research use only, not for use in diagnostic procedures.
Detailed Procedure of the Test
First, remove any extra micro ELISA plate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it. Add 100μL standard or sample to each well and incubate them covered for 2 hours at 37°C. Afterwards, remove the liquid and add 100μL Biotinylated Detection Ab to each well. Gently shake the plate to mix and incubate covered for 1 hour at 37°C. Next, aspirate and wash the plate thrice, then add 100μL HRP Conjugate to each well, cover and incubate for 30 minutes at 37°C. The aspiration/wash process is then repeated five times before adding 90μL Substrate Reagent. Incubate these covered for about 15 minutes at 37°C and then add 50 μL Stop Solution to each well. The color in the wells should change from blue to yellow.
Results Analysis
A microplate reader can be used to measure the optical density (OD) within 30 minutes at 450 nm wavelength. The OD value is proportional to the concentration of Human IL-8. You can calculate the concentration of Human IL-8 in the samples by comparing the OD of the samples to the standard curve.
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