Specificity
Alkaline phosphatase (AP). MAB1012 has a high affinity and recognizes an AP determinant resistant to denaturation by SDS-PAGE. It is therefore ideally suited for sensitive and specific detection of AP fusion proteins by Western blot analysis of E. coli transformants expressing fusion products.
Immunogen
Epitope: E. coli, bacterial only
F201C1B
Application
Anti-Alkaline Phosphatase Antibody, E. coli, bacterial only detects level of Alkaline Phosphatase & has been published & validated for use in IP, WB & IC.
Western blot at 1:5,000. 42-45kDa on SDS-PAGE,reducing gels for natural E. coli alkaline phosphatase monomer. Alkaline phosphatase fusion proteins will vary depending upon the target fusion protein.
Immunocytochemistry: reacts with E.coli. AP fusion protein targets in acetone fixed cell preparations. 1:4000, other fixatives or conditions untested.
ASSAY:
Preparation of E. coli TnphoA transformants: E. coli strain CC118 was transformed with plasmid pGEM-3Z containing TnphoA insertional mutations in the p101 gene of Mycoplasma hyorhinis, which encodes a protein with a typical N-terminal prokaryotic single peptide (Yogev et al. 1991).
Identification of fusion protein with MAB1012 transformants: Transformants are grown in 2XYT medium to OD600=0.6. Cells were centrifuged 3 minutes at 10,000 x g, suspended in SDS-PAGE sample buffer, heated at 100°C for 5 minutes, frozen and thawed and centrifuged as above at room temperature to remove insoluble material. The sample is applied at 9% to a SDS-PAGE gel, and Western immunoblot is performed as described (Yogev et al. 1991).
Immunoprecipitation: 5μL of antibody per 500μL of lysate in RIPA or 0.5% triton X-100 solutions.
Optimal working dilutions must be determined by end user.
Physical form
Ascites. Contains no preservative.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
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- UPC:
- 12352204
- Condition:
- New
- Availability:
- 3-5 Days
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- MPN:
- MAB1012
- Temperature Control Device:
- Yes
