Sandwich ELISA analysis of human GM-CSF was performed using a Human GM-CSF Colorimetric ELISA kit (Product # EHGMCSF) by loading 50 ul per well of a mouse anti-human GM-CSF biotinylated antibody (Product # M501B) at a concentration of 0.05 ug/ml and 50 ul of human GM-CSF Recombinant Protein (Product # SGMCSF) at 600, 240, 96, 38.4, 15.4, and 0 pg/ml (both in quadruplicate) across a 9 ug/ml mouse anti-human GM-CSF (Product # M500A) pre-coated plate. The plate was incubated for 3 hours at room temperature, washed, and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at 1:2000 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 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.
The human GM-CSF Standard is calibrated against NIBSC GM-CSF Reference Standard lot #88/646. 1 Thermo pg = 1 NIBSC pg = 0.01 NIBSC units.
Lyophilized product is shipped on cold packs. Stable until expiration if stored desiccated at -20°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.
1500 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.