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.
Filter by Application:
E ResetHuman anti Denosumab:HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD26295_hIgG1
- Isotype
- IgG1
- Specificity
- Denosumab
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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. |
- 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
- Buffer Solution
- Phosphate buffered saline
- 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 1.0 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 registered trademarks of Amgen Inc.
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 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
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Human anti Denosumab | HCA281 | 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 | ||||||
| Human anti Denosumab | HCA288 | E | 0.1 mg |
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| Description | Human anti Denosumab | ||||||
- Synonyms
- Prolia
- Xgeva
- 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
View more products with Denosumab specificity
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