|Product: ELISA MAX™ Standard Set Mouse IL-10|
|Catalog No.: 431412|
|Trim Lajqi, PhD student FSU Jena|
|Measurement of IL10|
|Overall:||Product Quality:||Ease of Use:|
|Measures the production of the anti-inflammatory IL10.|
|Application:||Measurement of IL10|
|Brief Protocol:||1. One day prior to running the ELISA, dilute Capture Antibody in 1X Coating Buffer A as described in Reagent Preparation. Add 100 ÃŽÂ¼L 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.
3. Wash plate 4 times with at least 300 ÃŽÂ¼L Wash Buffer per well and blot residual buffer by firmly tapping plate upside down on absorbent paper. All subsequent washes should be performed similarly.
4. To block non-specific binding and reduce background, add 200 ÃŽÂ¼L 1X Assay Diluent A 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 (if necessary) and standards. Wash plate 4 times with Wash Buffer.
8. Add 100 ÃŽÂ¼L/well of standards or samples to the appropriate wells. If dilution is required, samples should be diluted in 1X Assay Diluent A before adding to the wells.
9. Seal plate and incubate at RT for 2 hours with shaking.
10. Wash plate 4 times with Wash Buffer.
11. Add 100 ÃŽÂ¼L of diluted Detection Antibody solution to each well, seal plate and incubate at RT for 1 hour with shaking.
12. Wash plate 4 times with Wash Buffer.
13. Add 100 ÃŽÂ¼L of diluted Avidin-HRP solution to each well, seal plate and incubate at RT for 30 minutes with shaking.
14. Wash plate 5 times with Wash Buffer.
15. Add 100 ÃŽÂ¼L Substrate Solution.
16. Stop reaction by adding 100 ÃŽÂ¼L of Stop Solution to each well.
17. Read absorbance at 450 nm within 15 minutes.
|Results Summary:||The production of IL10 as anti-inflammatory cytokine.|
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