SYSY uses the following antigen retrieval buffers, which reflect the most popular buffers for HIER:
- Citrate Buffer (10 mM Citrate, 0.05% Tween 20, pH 6.0)
- Tris-EDTA buffer (10 mM Tris base, 1 mM EDTA, 0.05% Tween 20, pH 9.0) *
- EDTA buffer (1 mM EDTA, 0.05% Tween, pH 8.0) *
* This buffer works excellently for most antibodies and can highly improve sensitivity, but often results in high background
HIER is extremely sensitive to pH, time, temperature, and buffer composition. Generally, the higher the temperature, the shorter the heating time and vice versa. Specially formulated commercially available antigen retrieval buffers may be superior to SYSY buffers.
There is no universal method for antigen retrieval and no single HIER solution that is optimal for all antigens. Therefore, we recommend optimization of HIER using different time and pH / buffer conditions empirically for each antibody and each tissue type.