Ramucirumab antibody | AbD53018
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% BSA, the VEGFR2-His/ramucirumab complex was formed by adding 5µg/mL ramucirumab to the wells coated with VEGFR2-His. Detection was performed using Human Anti-Ramucirumab Antibody, clone AbD53018ad (TZA052) 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 ramucirumab, VEGFR2-His, human IgG1 kappa control or NHS at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, the VEGFR2-His/ramucirumab complex was formed by adding 5µg/mL ramucirumab to the wells coated with VEGFR2-His. Detection was performed using Human Anti-Ramucirumab Antibody, clone AbD53018ad (TZA052) titrated to the given concentrations in PBST followed by an HRP conjugated Anti-DYKDDDDK tag Antibody in HISPEC diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data for ramucirumab are shown as the mean of three measurements.
A microtiter plate was coated overnight with VEGFR2-His at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, a pre-incubated mixture of ramucirumab (0.15 µg/ml) plus increasing concentrations of Human Anti-Ramucirumab Antibody, clone AbD53018ad (TZA052) was added. The inhibitory Human Anti-Ramucirumab Antibody, clone AbD51673ad (TZA050) was used as a negative control. Free ramucirumab, still capable of binding to the VEGFR2-His coated plate was detected using HRP conjugated Mouse Anti-Human IgG (Fc) CH2 Domain Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Ramucirumab Antibody, clone AbD51673ad (TZA050) 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 ramucirumab. Detection was performed using HRP conjugated Human Anti-Ramucirumab Antibody, clone AbD53018pap (TZA052P) at a concentration of 0.5 µg/mL in HISPEC Assay Diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.
Human anti Ramucirumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD53018
- Isotype
- Fab antibody
- Specificity
- Ramucirumab
Antibody Quality and Performance Guarantee
TZA052
| Human Anti-Ramucirumab Antibody, clone AbD53018 is a non-paratope specific, non-inhibitory anti-idiotypic antibody (Type 2) that specifically recognizes the humanized monoclonal antibody drug ramucirumab. It recognizes free ramucirumab and ramucirumab in complex with its target, the human vascular endothelial growth factor receptor 2 (VEGFR2). It can be used to measure total levels of ramucirumab and biosimilars in patient samples. The monovalent Fab format of this antibody, clone AbD53018ad (TZA052) is tagged with a SpyTag2 at the C-terminus of the Fab heavy chain, enabling the user to couple it to a SpyCatcher reagent for conversion to alternative formats in less than one hour. This antibody is also available as a bivalent Fab antibody pre-coupled to the HRP-conjugated BiSpyCatcher2 (TZC002P) under product code TZA052P for direct detection of ramucirumab in bioanalysis assays. A pair of anti-ramucirumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure total drug. Clone AbD53018 can be used as a detection antibody to develop a pharmacokinetic (PK) bridging assay together with TZA050 as the capture antibody. Ramucirumab (trade name Cyramza) is a humanized monoclonal antibody of the IgG1/kappa isotype that binds to the extracellular VEGF-binding domain of VEGFR2 to prevent VEGF ligands to bind to VEGFR2. Ramucirumab therefore blocks the activation of VEGFR2 and the downstream angiogenesis pathways involved in tumor development and progression. Ramucirumab is indicated, alone or in combination with another drug for the treatment of advanced gastric adenocarcinoma and non-small cell lung cancer. |
- Product Form
- A monovalent human recombinant Fab (lambda light chain) selected from the HuCAL® phage display library, expressed in a proprietary E. coli. strain. The antibody is tagged with a DYKDDDDK tag, a SpyTag version 2 (VPTIVMVDAYKRYK) and a His6-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
- 0.0095% MIT
- Immunogen
- Ramucirumab
- Affinity
- The monovalent intrinsic affinity of AbD53018ad was measured as KD = 17 nM by real time, label free molecular interaction analysis on immobilized ramucirumab.
- 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.
His-tag is a registered trademark of EMD Biosciences.
Cyramza is a trademark of Eli Lilly. - 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 for coupling a Fab antibody with a SpyTag to a SpyCatcher can be found at SpyTag and SpyCatcher Products, and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- Clone AbD53018ad can be used in indirect ELISA to detect ramucirumab. It can also be used as a detection antibody to develop a pharmacokinetic (PK) bridging ELISA for ramucirumab, together with TZA050 as the capture reagent. Note that both TZA050 (capture) and TZA052 (detection) contain the same peptide tags. Therefore, using anti-tag antibodies for detecting TZA052 is not possible.
- Synonyms
- Cyramza
TZA052
168736
If you cannot find the batch/lot you are looking for please contact our technical support team for assistance.
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