Intended Use:The Bovine Interleukin-17A (IL-17A) ELISA Development Kit is for the quantitative determination of Bovine IL-17A in cell culture supernatants. 

Quantity: 2 Plate Set
Storage: 2-8ºC
Sample Type: Cell Culture Supernatants

Kit Contents

• Bovine IL-17A Coated Plate (2 plates)
• Bovine IL-17A Standard
• Bovine IL-17A Detection Antibody
• Streptavidin-HRP

ELISA Procedure:

1. Prepare Standard and cell culture supernatant sample dilutions in Standard and Sample Diluent.
2. Add 100 uL of Standard or sample to appropriate wells.
3. Note: Run each Standard or sample in duplicate.
4. Cover plate with Plate Sealer and incubate at room temperature (20-25C) for 1 hour.
5. Wash plate FOUR times with Wash Buffer.
6. Note: Gently squeeze the long sides of plate frame before washing to ensure all strips remain securely in the frame. Empty plate contents. Use a squirt wash bottle to vigorously fill each well completely with 1X Wash Buffer, then empty plate contents. Repeat procedure three additional times for a total of FOUR washes. Blot plate onto paper towels or other absorbent material.
7. Add 100 uL of Detection Antibody Working Solution to each well.
8. Cover plate with Plate Sealer and incubate at room temperature for 1 hour.
9. Wash plate FOUR times with Wash Buffer as described in step 4.
10. Add 100 uL of Streptavidin-HPR Working Solution to each well.
11. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.
12. Wash plate FOUR times with Wash Buffer as described in step 4.
13. Add 100 uL of TMB Substrate Solution to each well.
14. Develop the plate in the dark at room temperature for 30 minutes.
15. Note: Do NOT cover plate with Plate Sealer.
16. Stop reaction by adding 100 uL of Stop Solution to each well.
17. Measure absorbance on a plate reader at 450 nm

Additional Information: