MAB1012

Anti-Alkaline Phosphatase Antibody, E. coli, bacterial only

ascites fluid, Chemicon^®

• Synonym(s): AP, Alk. Phos.
• UNSPSC Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

Properties

• biological source: mouse
• Quality Level: 100
• antibody form: ascites fluid
• antibody product type: primary antibodies
• clone: monoclonal
• species reactivity: E. coli
• manufacturer/tradename: Chemicon^®
• technique(s): immunocytochemistry: suitable; immunoprecipitation (IP): suitable; western blot: suitable
• isotype: IgG2a
• UniProt accession no.: P00634
• shipped in: dry ice
• target post-translational modification: unmodified
• Gene Information: Escherichia coli ... PhoA(945041)

Description

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

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable