Vedolizumab antibody | AbD30136
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-Vedolizumab Antibody, clone AbD30136 (HCA292) 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 vedolizumab, human IgG1/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-Vedolizumab Antibody, clone AbD30136 (HCA292) titrated to the given concentrations in PBST, followed by detection with a Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Vedolizumab Antibody, clone AbD30136 (HCA292) 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 vedolizumab. Detection was performed using HRP conjugated Human Anti-Vedolizumab Antibody, clone AbD30136_hIgG1 (HCA293) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P).
A microtiter plate was coated overnight with Human Anti-Vedolizumab Antibody, clone AbD30136 (HCA292) 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 vedolizumab. Detection was performed using HRP conjugated Human Anti-Vedolizumab Antibody, clone AbD30138_hIgG1 (HCA294) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Vedolizumab Antibody, clone AbD30136 (HCA292) 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 vedolizumab. Detection was performed using HRP conjugated Human Anti-Vedolizumab Antibody, clone AbD30139_hIgG1 (HCA295) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P).
A microtiter plate was coated overnight with recombinant human α4β7 integrin at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of vedolizumab (0.3 µg/ml), spiked with increasing concentrations of Human Anti-Vedolizumab Antibody, clone AbD30136 (HCA292) was added. Free vedolizumab, still capable of binding to the human α4β7 integrin coated plate, was detected using a Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human anti Vedolizumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD30136
- Isotype
- HuCAL Fab monovalent
- Specificity
- Vedolizumab
Antibody Quality and Performance Guarantee
| Human Anti-Vedolizumab Antibody, clone AbD30136 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug vedolizumab. The antibody does not recognize free recombinant human α4β7 integrin or vedolizumab in complex with α4β7 integrin and can be used to measure free vedolizumab levels in serum from patients. A pair of anti-vedolizumab 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-vedolizumab antibody in full immunoglobulin format, clone AbD30138_hIgG1 (HCA294) as the detection antibody. Vedolizumab (Entyvio) is a humanized antibody (IgG1/kappa), derived from a murine antibody. It is used for the treatment of both moderate to severe ulcerative colitis and moderate to severe Crohn's disease, primarily in patients who have lost response to, or are intolerant to inhibitors of tumor necrosis factor alpha (TNFα), such as infliximab, adalimumab. The drug binds specifically to human α4β7 integrin, a key mediator of gastrointestinal inflammation expressed on the surfaces of T and B lymphocytes. By selectively inhibiting the binding of α4β7 integrin to the mucosal addressin cell adhesion molecule-1 (MAdCAM-1), vedolizumab prevents leukocyte binding to the endothelial surface and extravasation into affected tissue. Specifically inhibiting this pathway alleviates gastrointestinal inflammation without impairing systemic immune responses. View a summary of all anti-vedolizumab antibodies |
- Product Form
- A monovalent human recombinant Fab (lambda 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.
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline.
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Immunogen
- Vedolizumab
- Affinity
- The intrinsic affinity of the monovalent form of Human anti Vedolizumab antibody, clone AbD30136 is KD=0.1 nM as measured by real time, label-free molecular interaction analysis on immobilized Vedolizumab.
- Approx. Protein Concentrations
- Antibody 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.
Entyvio is a trademark of Millennium Pharmaceuticals, Inc.
His-tag is a trademark of EMD Biosciences.
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
- Human anti Vedolizumab antibody, clone AbD30136 can be used in a direct or indirect ELISA or as capture antibody for vedolizumab in a bridging ELISA together with (HCA293, HCA294, HCA294P, HCA295) as the detection reagent.
Protocol: PK bridging ELISA to measure free drug .
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Rat anti DYKDDDDK Tag | MCA4764 | E F IF IP P WB | 0.2 mg |
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| Description | Rat anti DYKDDDDK Tag | ||||||
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
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| Description | Mouse anti Penta Histidine Tag:HRP | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| HuCAL Fab-FH Negative Control | HCA045 | E | 0.1 mg |
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| Description | HuCAL Fab-FH Negative Control | ||||||
| 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 Vedolizumab | HCA293 | E | 0.1 mg |
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| Description | Human anti Vedolizumab | ||||||
| Human anti Vedolizumab | HCA294 | E | 0.1 mg |
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| Description | Human anti Vedolizumab | ||||||
| Human anti Vedolizumab:HRP | HCA294P | E | 0.1 mg | Log in | |||
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| Description | Human anti Vedolizumab:HRP | ||||||
| Human anti Vedolizumab | HCA295 | E | 0.1 mg |
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| Description | Human anti Vedolizumab | ||||||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | Log in | |||
| List Price | Your Price | ||||||
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| Description | LYNX Rapid HRP Antibody Conjugation Kit | ||||||
- Synonyms
- Entyvio
- 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.
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