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ProtoBlot II AP System with Stabilized Substrate, Human

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • Designed for rapid, sensitive alkaline phosphatase (AP) detection of proteins or other macromolecular antigens immobilized on nitrocellulose or PVDF membranes. Proteins can be transferred from gels after electrophoresis (Western blots) or bound directly from solution ("dot" blots). The Western Express™ Fast Blotting Protocol is included with the system and can reduce dramatically the time required to perform immunodetection, with detection of dot blots in 30–45 minutes and Western blots in 1–2 hours.

    The ProtoBlot® II AP Systems contain BSA as a stabilizer and 0.05% sodium azide as a preservative. In addition, the system contains Western Blue® Stabilized Substrate for AP, a ready-to-use solution of BCIP/NBT.

    For many applications, AP conjugates are superior to HRP conjugates:

    • Tenfold greater sensitivity of detection
    • Not inhibited by the presence of azide
    • Substrate is not prone to fading during long-term storage
    • Protocols provided for both PVDF and nitrocellulose membranes


    1. Towbin, H. et al. (1979) Proc. Natl. Acad. Sci. USA 76, 4350–4.
    2. Burnette, W.N. (1981) Anal. Biochem. 112, 195–203.
    3. Hawkes, R. et al. (1982) Anal. Biochem. 119, 142–7.
    4. Larson, G. (1992) Promega Notes 35, 18–20.
    5. Blake, M.S. et al. (1984) Anal. Biochem. 136, 175–9.
  • Protocols

    Complete Protocol

    Download PDF

    ProtoBlot® II AP System with Stabilized Substrate Technical Manual

    PDF (853 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    +2C TO +10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources