Denosumab antibody | AbD26296
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-Denosumab Antibody (clone AbD26296) at a concentration of 2 μg/ml and an HRP conjugated anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with denosumab, human IgG2/kappa or human IgG1/lambda at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Denosumab Antibody (clone AbD2629) titrated to the given concentrations in PBST, followed by detection with Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) and QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Denosumab Antibody (clone AbD26296) at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of denosumab. Detection was performed using Human Anti-Denosumab:HRP Antibody clone AbD26295_hIgG1 (HCA280P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of four measurements.
A microtiter plate was coated overnight with recombinant human RANKL at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, denosumab was added (0.3 μg/ml), spiked with increasing concentrations of each of three Human Anti-Denosumab Antibodies, clone AbD26295_hIgG (HCA280), clone AbD26862_hIgG1 (HCA282), both in monovalent Fab format in this assay, and clone AbD26296 (HCA288). Detection of denosumab was performed using Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.
Human anti Denosumab
Anti-denosumab antibody is a recombinant, inhibitory anti-idiotypic antibody in monovalent Fab format; it is recommended as capture antibody in a PK bridging ELISA format with HCA280 for detection.
- Product Type
- Monoclonal Antibody
- Clone
- AbD26296
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA288 | E | 0.1 mg |
|
Human Anti-Denosumab Antibody, clone AbD26296 is a paratope specific, anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug denosumab. The antibody does not recognize free RANKL (receptor activator of nuclear factor kappa-B ligand), or denosumab in complex with human RANKL and can be used to measure the levels of free denosumab and biosimilar products in bioanalytical assays.
A pair of anti-denosumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as the capture antibody, paired with an anti-denosumab antibody in full immunoglobulin format, clone AbD26295_hIgG1 (HCA280) as the detection antibody.
Denosumab (Prolia, Xgeva) is a fully human monoclonal antibody (IgG2/kappa) for the treatment of osteoporosis, treatment-induced bone loss, bone metastases, multiple myeloma, and giant cell tumor of bone. The drug specifically binds to human RANKL, a protein that acts as the primary signal to promote bone removal/resorption and prevents RANKL from activating its receptor, RANK, on the surface of osteoclasts and their precursors.
View a summary of all anti-denosumab antibodies.
A pair of anti-denosumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as the capture antibody, paired with an anti-denosumab antibody in full immunoglobulin format, clone AbD26295_hIgG1 (HCA280) as the detection antibody.
Denosumab (Prolia, Xgeva) is a fully human monoclonal antibody (IgG2/kappa) for the treatment of osteoporosis, treatment-induced bone loss, bone metastases, multiple myeloma, and giant cell tumor of bone. The drug specifically binds to human RANKL, a protein that acts as the primary signal to promote bone removal/resorption and prevents RANKL from activating its receptor, RANK, on the surface of osteoclasts and their precursors.
View a summary of all anti-denosumab antibodies.
Product Details
- Product Form
- A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL® phage display library, expressed in E. coli. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied as a liquid.
- Preparation
- Metal chelate affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Denosumab
- Affinity
- The monovalent intrinsic affinity of AbD26296 was measured as KD = 7.1 nM by real time, label free molecular interaction analysis on immobilized denosumab
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Guarantee
- 12 months from date of despatch
More Information
- 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. Prolia and Xgeva are registered trademarks of Amgen Inc.
His-tag is a registered trademark of EMD Biosciences. - Licensed Use
- For in vitro research purposes and for commercial applications for the provision of in vitro testing services to support preclinical and clinical drug development. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.
- Regulatory
- For research purposes only
Applications of Denosumab antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |
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
- ELISA
- Clone AbD26296 can be used in a direct or indirect ELISA system or as capture antibody for denosumab in a bridging ELISA together with HCA280P (AbD26295_hIgG1) as the detection reagent.
Protocol: PK bridging ELISA to measure total drug
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Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Denosumab | HCA280 | E | 0.1 mg |
|
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| Human anti Denosumab:HRP | HCA280P | E | 0.1 mg |
|
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| Human anti Denosumab | HCA281 | E | 0.1 mg |
|
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| Human anti Denosumab | HCA282 | E | 0.1 mg |
|
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| LYNX Rapid HRP Antibody Conjugation Kit | LNK001P | CJ | 1 Conjugation For 400µg Antibody | ||
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
|
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