Denosumab antibody | AbD26295_hIgG1
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 HRP conjugated Human Anti-Denosumab Antibody, clone AbD26295_hIgG1 (HCA280P) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and 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 AbD26295 (here as his-tagged, monovalent Fab fragment) 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 (HCA288) 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 denosumab 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 Human Anti-Denosumab Antibody, clone AbD26295_hIgG1 (HCA280, black line; KD = 1.8 nM), clone AbD26296_hIgG1 (HCA281,red line; KD = 7.1 nM) or clone AbD26862_hIgG1 (HCA282, blue line; KD = 0.8 nM). Detection was performed using HRP conjugated denosumab followed by QuantaBlu Fluorogenic Peroxidase. HRP conjugation of the denosumab used in this assay was performed using a LYNX Rapid HRP Conjugation Kit®. Data is shown as the mean of three 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_hIgG1 (HCA280), and 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:HRP
Anti-denosumab antibody is a recombinant, inhibitory, anti-idiotypic antibody in IgG1 format for use in bioanalytical assays to measure denosumab and biosimilars. It can be used as detection antibody in a PK bridging ELISA with HCA288, and as reference standard in an ADA bridging ELISA.
Human anti Denosumab
Anti-denosumab antibody is a recombinant, inhibitory, anti-idiotypic antibody in IgG1 format for use in bioanalytical assays to measure denosumab and biosimilars. It can be used as detection antibody in a PK bridging ELISA with HCA288, and as reference standard in an ADA bridging ELISA.
- Product Type
- Monoclonal Antibody
- Clone
- AbD26295_hIgG1
- Isotype
- IgG1
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA280P | E | 0.1 mg |
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| HCA280 | E | 0.1 mg |
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Human Anti-Denosumab Antibody, clone AbD26295_hIgG1 is a paratope specific anti-idiotypic antibody that specifically recognizes the free human monoclonal antibody 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 free denosumab and biosimilar products.
Clone AbD26295_hIgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay.
A pair of anti denosumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in full immunoglobulin format, is recommended as the detection antibody paired with clone AbD26296 (HCA288) in monovalent Fab format as the capture 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.
Clone AbD26295_hIgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay.
A pair of anti denosumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in full immunoglobulin format, is recommended as the detection antibody paired with clone AbD26296 (HCA288) in monovalent Fab format as the capture 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
- Human IgG1 antibody (kappa light chain) selected from the HuCAL® phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) – liquid
- Product Form
- Human IgG1 antibody (kappa light chain) selected from the HuCAL® phage display library and expressed in a human cell line - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Denosumab
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=1.8 nM by real time, label-free molecular interaction analysis on immobilized denosumab
- Approx. Protein Concentrations
- IgG concentration 0.1 mg/ml
- Approx. Protein Concentrations
- IgG concentration 0.5 mg/ml
Storage Information
- Storage
- Store at -70oC.
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. - 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
- 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.
- 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.
- 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 | |||
| 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.
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
- Technical Advice
- Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- This product may be used in a direct ELISA or as a detection antibody in a bridging ELISA together with HCA288 (AbD26296) as the capture reagent.
Protocol: PK bridging ELISA to measure total drug - ELISA
- This product may be used in direct or indirect ELISA and in an ELISA bridging assay as a detection antibody together, when conjugated to HRP, with HCA288 as the capture reagent. It is also suitable as a fully human control or calibrator antibody in an ADA bridging ELISA.
Protocol: PK bridging ELISA to measure free drug
Protocol: ADA bridging ELISA
Copyright © 2020 Bio-Rad Antibodies (formerly AbD Serotec)
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Human anti Denosumab | HCA281 | E | 0.1 mg |
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| Human anti Denosumab | HCA282 | E | 0.1 mg |
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| Human anti Denosumab | HCA288 | E | 0.1 mg |
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| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| 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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| Human anti Denosumab | HCA288 | E | 0.1 mg |
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| LYNX Rapid HRP Antibody Conjugation Kit | LNK001P | CJ | 1 Conjugation For 400µg Antibody |
