Casein, A2 beta Antibody

A synthetic peptide (PGPIPNSLP, aa: 78-86) conjugated to KLH has been used as immunogen. Bovine A1 beta casein differs from bovine A2 beta casein by one amino acid (H82 -> P82)

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.