Product: ELISA MAX™ Standard Set Mouse IL-6 |
Catalog No.: 431301 |
Julia Dickow, M.Sc. Essen University Hospital |
Mouse IL-6 ELISA MAX™ Standard | |||
Overall: | Product Quality: | Ease of Use: | |
Good working ELISA for quantification of cytokine production. |
Experimental Design |
Application: | ELISA |
Cells used: | In vitro cultured CD8 T cells with DCs |
Brief Protocol: | Dilute capture antibody 1:200 in 1 x coating buffer and incubate over night at 4 °C. Next day, wash plate 4 times with 200 µl of 1 x Wash Buffer (PBS + 0,05 % Tween-20). Block plate by adding 200 µl of Assay Diluent (PBS + 10 % FCS, sterile filtrated) for 1hour. Wash plate 4 times. Add diluted standards, as well as (serial-diluted) samples and incubate for 2 hours. Wash plate 4 times. Add 200 µl of diluted detection antibody (1:200 in Assay Diluent) and incubate for 1 hour. Wash plate 4 times. Add 200 µl of diluted Avidin-Antibody (1:1000 in Assay Diluent) and incubate for 30 minutes. Wash plate 5 times. Add 100 µl of TMB Substrate, incubate for 20 minutes and then stop the reaction with 2 N H2SO4. Measure absorbance at 570 nm (reference filter 450 nm). |
Results Summary: | Suitable for measurement of produced IL-6 by cells cultured in vitro. |
Additional Notes: |
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