Prepare Immunogenfrom SDS-PAGE Gel

For antibody production, you may use SDS-PAGE to separate the protein of interest from others. The following is a guideline for preparation of immunogen from SDS-PAGE.

Procedure

• Step 1 Prepare up to 2 mg total protein sample for loading.
• Step 2 Prepare aMinigelor standard SDS-PAGE gel using 1 mm thickness spacers and a prep-comb (with 1 big flat tooth) to make a wide flat well, deep enough to meet the loading volume.
• Step 3 Run the gel as regular SDS-PAGE procedure to separate the protein of interest.
• Step 4 Stain the gel withcoomassieblue. Destainthe gel and identify the protein band of interest.
• Step 5 Cut the band of interest using the whole width of the gel .
• Step 6 Transfer the gel strip with protein band into a tube, and send the sample for antibody production. The sample can be sent at room temperature.