Sandwich ELISA analysis of mouse TNF-alpha was performed using a Mouse TNF-alpha Colorimetric ELISA kit (Product # EMTNFA) by loading 50 ul per well of Mouse TNF-alpha Recombinant Protein (Product # SMTNFA) in dodecuplicate at 2450, 350, 50, and 0 pg/ml across a 3 ug/ml rat anti-mouse TNF-alpha (Product # MM350C ) pre-coated plate and incubating for 2 hours at room temperature along with 50 ul per well of rat anti-mouse TNF-alpha biotinylated antibody (Product # MM350DB). The plate was washed and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at 1:30,000 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.
This Mouse TNFalpha Standard is calibrated against NIBSC Mouse TNFalpha Reference Standard lot #88/532. 1 Thermo pg = 0.71 NIBSC pg = 0.14 NIBSC unit.
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.
2450 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.