MAB1012

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

ascites fluid, Chemicon^®

• Synonym(s): AP, Alk. Phos.
• UNSPSC Code: 12352203
• NACRES: NA.41
• eCl@ss: 32160702
• Conjugate: unconjugated
• Clone: monoclonal
• Application: ICC, IP, WB
• Citations: 13

Properties

• biological source: mouse
• conjugate: unconjugated
• 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

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.

Biochem/physiol Actions

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.

Physical form

Ascites. Contains no preservative.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety Information

• Storage Class: 10 - Combustible liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable