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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| E. coli | IP, WB, ICC | M | Ascites | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB1012 |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-Alkaline Phosphatase Antibody, E. coli, bacterial only |
| Alternate Names |
|
| Product Information | |
|---|---|
| Format | Ascites |
| Presentation | Ascites. Contains no preservative. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Alkaline Phosphatase Antibody, E. coli, bacterial only detects level of Alkaline Phosphatase & has been published & validated for use in IP, WB & IC. |
| Key Applications |
|
| Application Notes | 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. |
| Biological Information | |
|---|---|
| Immunogen | F201C1B |
| Epitope | E. coli, bacterial only |
| Host | Mouse |
| 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. |
| Isotype | IgG2a |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | Alkaline phosphatases are non-specific phosphomonoesterases that catalyze the hydrolysis reaction via a phosphoseryl intermediate to produce inorganic phosphate and the corresponding alcohol, optimally at high pH. Alkaline phosphatase exists as a dimer, each monomer binding 2 zinc atoms and one magnesium atom, which are essential for enzymatic activity. |
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P00634# alkaline phosphatase activity COFACTOR: Binds 1 magnesium ion; Binds 2 zinc ions. SIZE: 471 amino acids; 49,439 Da SUBUNIT: Homodimer. SUBCELLULAR LOCATION: Periplasm, Periplasmic space. PTM: Disulfide bond; Phosphoprotein. SIMILARITY: SwissProt: P00634 ## Belongs to the alkaline phosphatase family. |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Store at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze/thaw cycles. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |