Human IL-8 ELISA Kit (High Sensitivity) is a 3h 30m sandwich Enzyme-Linked Immunosorbent Assay (sELISA) optimized for the detection of low -abundance targets, designed for the in vitro quantitative determination of human IL-8 in serum, plasma, and other biological fluids.
Assay Type
Sandwich (quantitative)
Principle of Assay
Human IL-8 ELISA Kit (A323171) employs the sandwich Enzyme-Linked Immunoassay technique for the quantitative measurement of low-abundance human IL-8 in serum, plasma, and 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. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Avidin conjugate is 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
Colorimetric Detection (OD450)
Instrument
Colorimetric Microplate Reader
Sample Type
Serum, plasma, and other biological fluids.
Sensitivity
0.29 pg/mL
Product Range
0.47-30 pg/mL
Assay Time
3h 30m
Reactivity
Human
Cross Reactivity
Specific for IL-8 with no reported cross-reactivity with other analogs or variants.
Recovery
Sample Type
n
Range
Average
Serum
8
91 - 106%
99%
EDTA Plasma
8
93 - 106%
99%
Cell Culture Media
8
93 - 108%
99%
Storage
Shipped at +4°C. Store kit components at +4°C or -20°C as instructed in the Protocol Manual. Do not use past expiration date! Avoid freeze/thaw cycles.
Linearity assessment of the Human IL-8 ELISA Kit (High Sensitivity) (A323171) across serial sample dilutions. Linearity was evaluated using Serum, EDTA Plasma, and Cell Culture Media samples serially diluted from 1:2 to 1:16. Recovery (%) was calculated relative to the expected Human IL-8 concentration at each dilution. Data are presented as mean recovery values from 8 replicates (n = 8).
Workflow of the Antibodies.com High Sensitivity ELISA Kit assay procedure. Samples or standards are added to antibody pre-coated wells and incubated to allow target binding. Following removal of unbound material, a biotinylated detection antibody is added, followed by HRP-conjugated streptavidin. After washing steps, TMB substrate is introduced to generate a colorimetric signal proportional to target concentration. The reaction is terminated with stop solution, and absorbance is measured at 450 nm for quantitative analysis.
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