Sandwich ELISA analysis of human IL-8 was performed using a Human IL-8 Colorimetric ELISA kit (Product # EH2IL8) by loading 50 ul per well of Human IL-8 Recombinant Protein (Product # SIL8) in dodecuplicate at 1000, 400, 160, 64, 25.6, and 0 pg/ml across a 3 ug/ml mouse anti-Human IL-8 (Product # M801) pre-coated plate and incubating for 1 hour at room temperature. The plate was washed, and then incubated with 50 ul per well of a biotinylated mouse anti-human IL-8 monoclonal antibody (Product # M802B) in duplicate at 0.25 ug/ml for 1 hour at room temperature. The plate was washed and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at a 1:4,000 dilution for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
|Tested species reactivity||Not Applicable|
|Host / Isotype||E.coli|
|Storage buffer||4% BSA|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Control (Ctrl)||Assay Dependent|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Calibrated to an in-house standard preparation.
Lyophilized product is shipped on cold packs. Stable until expiration if stored desiccated at 2-8°C. Reconstituted cytokine standards should be stored in aliquots at -70°C. Each aliquot should be used only once and discarded afterwards. Stability of reconstituted and frozen cytokine standards may vary. Controls should be run with each standard curve.
Sufficient For: ELISA standard curve with replicates or as a control in Western blots.
2500 pg/ml when reconstituted at the recommend volume.
Reconstitute with distilled water at volume indicated on label. Use or freeze product within 1 hour of reconstitution.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.