Ipilimumab antibody | AbD34294
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-Ipilimumab Antibody, clone AbD34294 (HCA331) at a concentration of 2 μg/ml and an HRP labeled anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with recombinant human CTLA-4, ipilimumab, human IgG1 kappa or human IgG1 lambda at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, the CTLA-4/ipilimumab complex was formed by adding 2 µg/ml ipilimumab to the wells coated with CTLA-4. Detection was performed using Human Anti-Ipilimumab Antibody, clone AbD34294 (HCA331) titrated to the given concentrations in PBST followed by an HRP conjugated anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with human CTLA-4 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 ipilimumab. Detection was performed using Human Anti-Ipilimumab antibody, clone AbD34294 (HCA331) at a concentration of 2 µg/ml followed by an HRP conjugated anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human Anti-Ipilimumab Antibody, clone AbD34294 specifically recognizes the monoclonal antibody drug ipilimumab when in complex with its target, CTLA-4 (cytotoxic T-lymphocyte antigen-4). The antibody does not recognize free ipilimumab or unbound recombinant human CTLA-4. It can be used to measure levels of ipilimumab in patient samples.
Clone AbD34294 is a monovalent Fab format antibody that can be used to develop a pharmacokinetic (PK) antigen capture assay to measure free ipilimumab captured via immobilized CTLA-4.
Ipilimumab (Yervoy) is a human IgG1/kappa antibody developed from a transgenic mouse. It has been approved by the FDA for the treatment of metastatic melanoma, renal cell carcinoma, and in combination with nivolumab for the treatment of previously treated microsatellite instability-high/deficient mismatch repair (MSI-H/dMMR) metastatic colorectal cancer. Ipilimumab activates the immune system by targeting CTLA-4, a protein receptor that downregulates the immune system. The action of cytotoxic T lymphocytes (CTLs) to recognize and destroy cancer cells is subject to an inhibitory mechanism that interrupts this destruction. Ipilimumab turns off this inhibitory mechanism and allows CTLs to function.
View a summary of all anti-ipilimumab antibodies
Clone AbD34294 is a monovalent Fab format antibody that can be used to develop a pharmacokinetic (PK) antigen capture assay to measure free ipilimumab captured via immobilized CTLA-4.
Ipilimumab (Yervoy) is a human IgG1/kappa antibody developed from a transgenic mouse. It has been approved by the FDA for the treatment of metastatic melanoma, renal cell carcinoma, and in combination with nivolumab for the treatment of previously treated microsatellite instability-high/deficient mismatch repair (MSI-H/dMMR) metastatic colorectal cancer. Ipilimumab activates the immune system by targeting CTLA-4, a protein receptor that downregulates the immune system. The action of cytotoxic T lymphocytes (CTLs) to recognize and destroy cancer cells is subject to an inhibitory mechanism that interrupts this destruction. Ipilimumab turns off this inhibitory mechanism and allows CTLs to function.
View a summary of all anti-ipilimumab antibodies
Product Details
- 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 double extended Strep-tag (SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK). This antibody is supplied as a liquid.
- Preparation
- StrepTactin affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Ipilimumab
- Affinity
- The monovalent intrinsic affinity of AbD34294 was measured as KD = 252 nM by real time, label free molecular interaction analysis on immobilized CTLA-4/ipilimumab complex.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Guarantee
- 12 months from date of despatch
More Information
- Acknowledgements
- Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
Yervoy is a trademark of Bristol-Myers Squibb Company. - 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.
- Regulatory
- For research purposes only
Applications of Ipilimumab antibody
| 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
- Clone AbD34294 is recommended for use as the detection reagent in an antigen capture assay to measure free ipilimumab captured via immobilized CTLA-4.
Protocol: PK antigen capture ELISA to measure bound drug exclusively
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Secondary Antibodies Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Rat anti DYKDDDDK Tag:HRP | MCA4764P | E P WB | 0.1 mg |
|
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Ipilimumab | HCA327 | E | 0.1 mg |
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| Human anti Ipilimumab | HCA328 | E | 0.1 mg |
|
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| Human anti Ipilimumab | HCA329 | E | 0.1 mg |
|
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| Human anti Ipilimumab | HCA330 | E | 0.1 mg |
|
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| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody |
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