Alkaline Phosphatase antibody | AbD25296_hIgG2
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Bacterial Alkaline Phosphatase:HRP Antibody (clone AbD25296_hIgG2) at a concentration of 2 μg/ml followed by Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
Western blot analysis was performed by loading 25 μl Hela lysate (lanes 2 and 3) and Precision Plus Protein™ Prestained Standards (lane 1) onto a AnykD™ Criterion™ TGX Stain-Free™ Protein Gel. The gel was run under reducing conditions and proteins were transferred to a nitrocellulose membrane. Lane 2 was probed with Anti-HSPA8 Antibody, clone AbD28028 Fab-A-FH format, at a concentration of 10 μg/ml. Lanes 2 and 3 were then probed with Human Anti-Bacterial Alkaline Phosphatase:HRP (clone AbD25296_hIgG2, HCA275P) at dilution of 1/1000. Visualization was carried out using the ChemiDoc™ MP Imaging System with an auto exposure. The Western blot bands (red) and protein standards (green) have been pseudocolored. Fab-A-FH format is a bivalent bacterial alkaline phosphatase fusion antibody (DYKDDDDK- and His-6-tags).
A microtiter plate was coated overnight with recombinant GFP antibody (His-tagged) at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using primary human anti-GFP antibodies in different bivalent Fab formats (e.g. Fab-A-V5Sx2) titrated to the given concentrations in PBST, followed by Human Anti-Bacterial Alkaline Phosphatase:HRP Antibody (clone AbD25296_hIgG2) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of triplicate measurements.
Human anti Bacterial Alkaline Phosphatase:HRP
This HRP conjugated Anti-Bacterial Alkaline Phosphatase Antibody is recommended for use as a secondary detection reagent with HuCAL antibodies in Fab-A and Fab-Max formats.
- Product Type
- Monoclonal Antibody
- Clone
- AbD25296_hIgG2
- Isotype
- IgG2
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA275P | E WB | 0.1 mg |
|
Human Anti-Bacterial Alkaline Phosphatase Antibody, clone AbD25296_hIgG2 recognizes bacterial alkaline phosphatase (BAP), a periplasmic enzyme which catalyzes the release of 5'- and 3'-phosphate groups from DNA, RNA and nucleotides. BAP can also dephosphorylate a number of proteins and alkaloids, and has an optimum pH of 8.0. BAP is encoded by the phoA gene, and forms a homodimer of two identical ~47 kDa subunits.
Fab-A and Fab-Max format antibodies generated using HuCAL® technology are formed via dimerization of BAP. The Anti-BAP Antibody HCA275P can be used as a secondary detection reagent with HuCAL antibodies in Fab-A and Fab-Max formats (Fab-Max is a modified BAP fusion antibody with inactivated enzymatic activity).
View further information about HuCAL antibody formats.
Fab-A and Fab-Max format antibodies generated using HuCAL® technology are formed via dimerization of BAP. The Anti-BAP Antibody HCA275P can be used as a secondary detection reagent with HuCAL antibodies in Fab-A and Fab-Max formats (Fab-Max is a modified BAP fusion antibody with inactivated enzymatic activity).
View further information about HuCAL antibody formats.
Product Details
- Target Species
- Bacterial
- Product Form
- Human IgG2 antibody (lambda light chain) selected from the HuCAL® phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
HRP Stabiliser (BUF052A) - Immunogen
- Bacterial Alkaline Phosphatase
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=38pM by real time, label-free molecular interaction analysis on immobilized HuCAL antibody in the Fab-A-FH format
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
Storage Information
- Storage
- Store at +4oC. DO NOT FREEZE
This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use - Guarantee
- 12 months from date of despatch
More Information
- UniProt
- P00634 Related reagents
- GO Terms
- GO:0005515 protein binding
- GO:0000287 magnesium ion binding
- GO:0004035 alkaline phosphatase activity
- GO:0008270 zinc ion binding
- GO:0030613 oxidoreductase activity, acting on phosphorus or arsenic in donors
- GO:0033748 hydrogenase (acceptor) activity
- GO:0042597 periplasmic space
- Acknowledgements
- Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
- Regulatory
- For research purposes only
Applications of Alkaline Phosphatase antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA | |||
| Western Blotting | 1/1,000 | 1/10,000 |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Technical Advice
- Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
Copyright © 2019 Bio-Rad Antibodies (formerly AbD Serotec)
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml |
Product Specific References
References for Alkaline Phosphatase antibody
-
Harth, S. et al. (2019) Generation by phage display and characterization of drug-target complex-specific antibodies for pharmacokinetic analysis of biotherapeutics.
MAbs. 11 (1): 178-90.
