Denosumab antibody | AbD26781_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 Human Anti-Denosumab:HRP Antibody clone AbD26781_hIgG1 (HCA283P) 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 RANKL, human IgG1/kappa, human IgG2/kappa or human IgG1/lambda at a concentration of 5 µg/ml. For complex formation, denosumab, at a concentration of 2µg/mL, was added to RANKL coated wells after blocking. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Denosumab:HRP Antibody clone AbD26781_hIgG1 (HCA283P) titrated to the given concentrations in PBST, followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data for binding to denosumab and to the RANKL-denosumab complex 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 four Human Anti-Denosumab Antibodies, clone AbD26295_hIgG1 (HCA280), clone AbD26862_hIgG1 (HCA282), clone AbD26781_hIgG1 (HCA283) all here in monovalent Fab format, 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. Note, although HCA283 is a non-inhibitory antibody, it shows a slight decrease in signal in this assay. This is most likely due to different binding modes of the denosumab-RANKL complex at higher denosumab concentrations (Arthur et al., 2012).
A microtiter plate was coated overnight with one of two Anti-Denosumab Antibodies, either clone AbD26295_hIgG1 (HCA280) or clone AbD26862_hIgG1 (HCA282) 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 AbD26781_hIgG1 (HCA283P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.
A microtiter plate was coated overnight with human RANKL at a concentration of 5 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 AbD26781_hIgG1 (HCA283P) in HISPEC Assay Diluent (BUF049A) at a concentration of 2 μg/ml, followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is presented as the mean of three measurements.
Human anti Denosumab:HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD26781_hIgG1
- Isotype
- IgG1
- Specificity
- Denosumab
Antibody Quality and Performance Guarantee
| Human Anti-Denosumab Antibody, clone AbD26781_hIgG1 is a non-inhibitory anti-idiotypic antibody that specifically recognizes free human monoclonal antibody drug denosumab, and the drug in complex with its target, RANKL (receptor activator of nuclear factor kappa-B ligand). The antibody does not recognize free RANKL and does not inhibit binding of denosumab to RANKL. A pair of anti-denosumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay. This antibody clone AbD26781_hIgG1 is recommended as the detection antibody, paired with either clone AbD26295_hIgG1 (HCA280) or clone AbD26862_hIgG1 (HCA282) as capture antibody. It can also be used as the detection antibody in an antigen capture format assay. This antibody is not suitable for the development of an anti-drug antibody (ADA) assay. 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 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
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Source
- HKB-11
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Denosumab
- Affinity
- The intrinsic affinity of the monovalent form of this antibody is KD=19 nM as measured by real time, label-free molecular interaction analysis on immobilized denosumab, and as KD =17 nM on the RANKL/drug complex.
- Approx. Protein Concentrations
- IgG concentration 0.1 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
Prolia and Xgeva are 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.
This product is shipped frozen.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| 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
- This antibody can be used as a detection antibody in a bridging ELISA together with HCA280 (clone AbD26295_hIgG1) or HCA282 (clone AbD26862_hIgG1) as capture reagents. This antibody can be used in an antigen capture ELISA. Protocol: PK bridging ELISA to measure free drug.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Denosumab | HCA280 | E | 0.1 mg |
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| Description | Human anti Denosumab | ||||||
| Human anti Denosumab | HCA282 | E | 0.1 mg |
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| Description | Human anti Denosumab | ||||||
References for Denosumab antibody
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Arthur, K.K. et al. (2012) In vitro stoichiometry of complexes between the soluble RANK ligand and the monoclonal antibody denosumab.
Biochemistry. 51 (3): 795-806.
- Synonyms
- Prolia
- Xgeva
HCA283P
153108
153716
1807
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