Omalizumab antibody | AbD20669
A microtiter plate was coated over night with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti omalizumab (clone AbD20669) at a concentration of 2 μg/ml, followed by an HRP-labelled anti-His tag in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate.
A microtiter plate was coated over night with Human IgG1/kappa, Human IgG1/lambda, trastuzumab or omalizumab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti omalizumab (clone AbD20669) titrated to the given concentrations in PBST, followed by an HRP-labelled anti-His tag in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate.
A microtiter plate was coated over night with Human anti omalizumab (clone AbD20669) 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 omalizumab. Detection was performed using HRP-conjugated Human anti omalizumab (clone AbD20669) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit®
A microtiter plate was coated over night with Human anti omalizumab (clone AbD20669) 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 omalizumab. Detection was performed using HRP-conjugated Human anti omalizumab (clone AbD20669_hIgG1, HCA235) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit.
A microtiter plate was coated over night with Human IgE (HCA171) at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of omalizumab (0.3 μg/ml) plus increasing concentrations of Human anti omalizumab (clone AbD20669) was added. Free omalizumab, still capable of binding to the IgE-coated plate, was detected using Goat anti Human IgG (Fc):HRP antibody and QuantaBlu fluorogenic peroxidase substrate.
Human anti Omalizumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD20669
- Isotype
- HuCAL Fab monovalent
- Specificity
- Omalizumab
Antibody Quality and Performance Guarantee
| Omalizumab (brand name Xolair) is a recombinant DNA-derived humanized IgG1 kappa monoclonal antibody used in the treatment of patients with moderate or severe asthma who have demonstrated a positive allergy skin test and whose symptoms are not controlled by inhaled corticosteroids. Allergic asthma is mediated by IgE released by B cells in response to allergen. Circulating IgE binds to the high-affinity IgE Fc receptor (FcεRI) expressed on basophils and mast cells, triggering the release of histamine, leukotrienes and other mediators associated with the pathophysiology of asthma. Omalizumab is directed against the Fc region of human immunoglobulin E (IgE). By binding to circulating IgE at the site of FcεRI binding, this therapeutic antibody prevents the interaction of IgE with its receptor thus limiting mediator release. Treatment with omalizumab has also been demonstrated to reduce the expression of FcεRI on mast cells and basophils, providing additional clinical benefit. Human anti omalizumab, clone AbD20669 is a paratope specific anti-idiotypic (Type 1) antibody that binds specifically to omalizumab but not to free IgE or the drug/immunoglobulin complex. This antibody can be used to measure omalizumab levels in serum from patients. Clone AbD20669 is particularly suited for use as a capture reagent in Pharmacokinetic (PK) assay development. Other Type 1 (HCA235) and Type 3 (HCA237, HCA238) antibodies to omalizumab are also available from Bio-Rad in IgG1 and Fab formats. This range of anti omalizumab antibodies forms part of a wider and expanding portfolio of anti-idiotypic antibodies that have been developed for the detection of biotherapeutics. |
|
- Product Form
- A monovalent human recombinant Fab 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.
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline.
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Immunogen
- Omalizumab.
- Affinity
- The intrinsic affinity of the monovalent form of Human anti Omalizumab antibody, clone AbD20669 is Kd = 1.1 nM as measured by real time, label free molecular interaction analysis on immobilized omalizumab.
- Approx. Protein Concentrations
- IgG concentration 0.5 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.
Xolair is a trademark of Novartis.
His-tag is a 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.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
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 as a capture reagent in PK assay development together with HCA235P as the detection reagent.
Protocol: PK bridging ELISA to measure free drug.
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HCA236
151574
155749
161493
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