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Sandwich ELISA analysis of human IL-1b was performed using a Human IL-1b Colorimetric ELISA Kit (Product # EH2IL1B) by loading 50 ul per well of Human IL-1b Recombinant Protein (Product # SIL1B) in quadruplicate at 400, 160, 64, 25.6, 10.24 and 0 pg/ml across a 5 ug/ml mouse anti-human IL-1b (Product # M421B) pre-coated plate and incubating for 3 hours at room temperature. The plate was washed then incubated with 50 ul per well of a biotinylated-mouse anti-human IL-1b monoclonal antibody (Product # M420BB) in quadruplicate at 0.07 ug/ml for 3 hours at room temperature. The plate was washed and incubated with 100 ul per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at 1:3000 for 30 minutes at room temperature. Detection was performed using 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|
|Published species reactivity||Human|
|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.
|ELISA (ELISA)||See 1 publications below|
The human IL-1beta Standard is calibrated against WHO IL-1beta Reference Standard lot #86/522. 1 Thermo pg = 1 WHO pg = 0.1 WHO units.
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.
1000 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.
Effect of age on surface molecules and cytokine expression in human dendritic cells.
SIL1B was used in ELISA to investigate the immunosenescence of human dendritic cells
|Ciaramella A,Spalletta G,Bizzoni F,Salani F,Caltagirone C,Bossù P||Cellular immunology (269:82)||2011|