Product: ELISA MAX™ Standard Set Mouse IL-1a

Catalog No.: 433402

Lajqi Trim, PhD Student FSU Jena

Mouse IL-1α ELISA
Overall:	Product Quality:	Ease of Use:
Mouse IL-1α production

Experimental Design

Application:	Measurement of IL-1 alpha
Cells used:	BV2 cells
Brief Protocol:	A day prior to running the ELISA, dilute Capture Antibody in 1X Coating Buffer. Add 100 μL of this Capture Antibody solution to all wells of a 96-well plate provided in this set. Seal plate and incubate overnight (16-18 hrs) between 2°C and 8°C. 2. Bring all reagents to room temperature (RT) prior to use. It is strongly recommended that all standards and samples be run in duplicate or triplicate. 3. Wash plate 4 times with at least 300 μL Wash Buffer per well. 4. To block non-specific binding and reduce background, add 200 μL 1X Assay Diluent per well. 5. Seal plate and incubate at RT for 1 hour with shaking on a plate shaker. 6. While plate is being blocked, prepare the appropriate sample dilutions and standards. Wash plate 4 times with Wash Buffer. 8. Add 100 μL/well of standards or samples to the appropriate wells. 9. Incubate at RT for 2 hours with shaking. 10. Wash plate 4 times with Wash Buffer. 11. Add 100 μL of diluted Detection Antibody solution and incubate for 1 hour. 12. Wash plate 4 times with Wash Buffer. 13. Add 100 μL of diluted Avidin-HRP solution to each well. 14. Wash plate 5 times with Wash Buffer. This will help minimize background. 15. Add 100 μL Substrate Solution. 16. Stop reaction by adding 100 μL of Stop Solution to each well. 17. Read absorbance at 450 nm.
Results Summary:	Production of IL-1 alpha
Additional Notes:	Incubation overnight is preferred at 4 Celsius degrees.

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Publication Status This data has not been published.

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