Product: ELISA MAX™ Deluxe Set Human IFN-γ

Catalog No.: 430105

Anne Rissiek, Dr. Universitätsklinikum Hamburg-Eppendorf (UKE)

Very Good ELISA Kit for Detection of Human IFNg in Cell Culture Supernatants
Overall:	Product Quality:	Ease of Use:
Was able to detect human IFNg from cell culture supernatants.

Experimental Design

Application:	ELISA
Cells used:	T cell culture supernatants
Brief Protocol:	Day1 1. Coated plate 100 µl/well of Capture Ab in Coating Buffer. Seal plate. Overnight, 4 °C. Day2 Bring all reagents to RT 2. Wash 4x with washing buffer, 300 µl. Allowing time for soaking ca 1 min during each wash step increases the effectiveness of the washes. Blot plate on absorbent paper to remove any residual buffer. 3. Block wells: 200 µl/well of 1x Assay Diluent. Seal plate and incubate 1 h at RT, while shaking. 4. Wash 4x with washing buffer. 5. Load plate Standard: 100 µl/well standard to appropriate wells. Samples: 100 µl/well samples to appropriate wells Seal the plate and incubate 2 h, RT, while shaking. 6. Wash 4x with Washing buffer. 7. Cytokine Detection: 100 µl/well Detection Ab. Seal plate and incubate 1 h at RT, while shaking. 8. Wash 4x with Washing buffer. 9. Detection Enzyme: 100 µl/well avidin-HRP. Seal plate and incubate 30 min at RT, while shaking. 10. Wash 5x with Washing buffer, soak each well 30 sec -1 min 11. Substrate Solution 100 µl/well TMB Substrate Solution. Seal plate and incubate 30 min, RT, in the dark --> blue 12. Stopping, add 100 µl/well Stop Solution --> yellow 13. Read plate at 450nm within 30 min.
Results Summary:	Was able to detect human IFNg from cell culture supernatants. Nice standard curve!
Additional Notes:

Data Image Click on the image for full size data.

Publication Status This data has been published. Pubmed

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