Product: ELISA MAX™ Deluxe Set Human IL-17A |
Catalog No.: 433915 |
Anne Rissiek, Dr. Universitätsklinikum Hamburg-Eppendorf (UKE) |
Human IL-17A ELISA | |||
Overall: | Product Quality: | Ease of Use: | |
Very good ELISA kit for detection of human IL-17A in cell culture supernatants |
Experimental Design |
Application: | ELISA |
Cells used: | Cell culture supernatants, human T cell culture |
Brief Protocol: | day1 1. Coate 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 1min 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, 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, RT, while shaking. 8. Wash 4x with Washing buffer. 9. Detection Enzyme, 100 µl/well avidin-HRP. Seal plate and incubate 30 min, RT, while shaking 10. Wash 5x with Washing buffer, soak each wlell 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 IL-17A from cell culture supernatants. Nice standard curve! |
Additional Notes: |
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