WB; ELISA Western blot and ELISA. Suggested working dilution for western blot is 1:1,000-1:5,000. The amount of milk per lane can be 0.05 µL-0.1 µL for Western blot. Sample Preparation: Milk should be diluted 1:10 in 0.1M NaOH. The reason for diluting (1:10) in 0.1M NaOH is because the milk protein is easier to dissolve in NaOH. The sample is then further diluted with PBS or other buffer and mixed at 1:1 ratio, to prepare for loading. For example, one can take the 1:10 dilution milk (0.5 µL-1 µL) and add into 9.5 µL or 9 µL PBS and mix with 10 µL SDS-PAGE Sample buffer, boil for 5 minutes, quick spin, and load on the gel. Recommended blocking buffer: TBS with 5% BSA. Recommended antibody dilution buffer: TBST containing 3% BSA. Biosensis recommends that optimal working dilutions should be determined by the end user.
免疫原
A synthetic peptide (PGPIHNSLP, aa: 78-86) conjugated to KLH has been used as immunogen. Bovine A2 beta casein differs from bovine A1 beta casein by one amino acid (P82 -> H82)
