ELISA protocol
Summary of Assay Procedure for Sandwich Kit:
-Prepare all reagents, samples and standards,
- Add 100 ul of standard or sample to each well,
-Seal the plate and incubate for 2 hrs at 37 °C
-Wash the plate 3 times with wash buffer solution,
-Add 100 ul of Biotin conjugated detection antibody,
-Seal the plate and incubate for 1 hr at 37 °C
-Wash plate 3 times with wash buffer solution,
-Add 100 ul of HRP conjugated detection antibody,
-Seal the plate and incubate for 30 mins at 37 °C
-Wash plate 5 times with wash buffer solution,
-Add 90 ul of TMB substrate solution,
-Seal the plate and incubate for 20 mins at 37 °C
-Add 50 ul of stop solution,
-Read the plate at 450 nm.
Summary of Assay Procedure for Competitive Kit:
-Prepare all reagents, samples and standards,
- Add 50 ul of standard or sample to each well,
-Immediately add 50 ul of Biotin conjugated detection antibody,
-Seal the plate and incubate for 1 hr at 37 °C
-Wash plate 3 times with wash buffer solution,
-Add 100 ul of HRP conjugated detection antibody,
-Seal the plate and incubate for 30 mins at 37 °C
-Wash plate 5 times with wash buffer solution,
-Add 90 ul of TMB substrate solution,
-Seal the plate and incubate for 20 mins at 37 °C
-Add 50 ul of stop solution,
-Read the plate at 450 nm.
Please note quantities of samples, reagents and incubation periods can vary between each kit, please refer to the product manual for further information.
