AffiELISA® OneStep® Human ANXA1 (Annexin A1) ELISA Kit

ELISA Assay Procedure 

1. Sample and Standard Preparation
   • Add 100μL of standard or sample to the wells.
   • Incubate for 90 minutes at 37°C.
2. Biotinylated Detection Antibody Application
   • Discard the liquid from the wells.
   • Immediately add 100μL of Biotinylated Detection Antibody (Ab) working solution to each well.
   • Incubate for 60 minutes at 37°C.
3. Washing
   • Aspirate the contents of the wells.
   • Wash the plate 3 times.
4. HRP Conjugate Application
   • Add 100μL of HRP (Horseradish Peroxidase) conjugate working solution to each well.
   • Incubate for 30 minutes at 37°C.
   • Aspirate and wash the plate 5 times.
5. Substrate Reaction
   • Add 90μL of Substrate Reagent to each well.
   • Incubate for 15 minutes at 37°C.
6. Stopping the Reaction
   • Add 50μL of Stop Solution to each well.
7. Measurement and Analysis
   • Read the optical density (OD) of each well at 450 nm using a spectrophotometer immediately after adding the Stop Solution.
   • Calculate the results based on the OD readings.

For more detailed information or specific instructions related to your Elisa OneStep kit, it's recommended to consult our technical support team.