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Sandwich ELISA analysis of human IL-5 was performed by coating wells of a 96-well plate with 110ul of a human IL-5 monoclonal antibody (Product # M551) in duplicate at 1, 2, 4, 5, 7, and 9ug/ml for at least 12 hours. The plate was blocked with a proprietary blocking solution for 1-4 hours at room temperature, and 50ul of mouse IL-5 recombinant protein (Product # SMIL5) was added in replicates of twelve at 400, 160, 64, 25.6, 10.24, and 0pg/ml for 1 hour at room temperature. The plate was washed and incubated 50ul of a biotinylated IL-5 monoclonal antibody (Product # M550B) was added to all test wells at 0.5ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of Streptavidin-HRP (Product # N504) at a 1:10,000 dilution for 30 minutes at room temperature. Detection was performed with 100ul per well of 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 450-550nm.
|Host / Isotype||unidentified baculovirus|
|Storage buffer||4% BSA|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|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.
International reference standard is not available. Calibrated to a Thermo reference standard.
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.
320 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.