Ipilimumab antibody | AbD34433
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% milk, detection was performed using Human Anti-Ipilimumab Antibody, clone AbD34433 (HCA330) 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 ipilimumab, human IgG1 kappa, human IgG1 lambda or hCTLA-4 at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using Human Anti-Ipilimumab Antibody, clone AbD34433 (HCA330) 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.
A microtiter plate was coated overnight with Human Anti-Ipilimumab Antibody, clone AbD34433 (HCA330) 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 ipilimumab. Detection was performed using HRP conjugated Human Anti-Ipilimumab Antibody, clone AbD34429ia (HCA329P) at a concentration of 2 µg/ml 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 recombinant human CTLA-4 at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, a pre-incubated mixture of HRP conjugated ipilimumab (0.3 µg/ml) plus increasing concentrations of Human Anti-Ipilimumab Antibody, clone AbD34433 (HCA330) in the monovalent Fab format was added. Free HRP conjugated ipilimumab, still capable of binding to the human CTLA-4 coated plate, was detected using QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human anti Ipilimumab
Human Anti-Ipilimumab Antibody is a recombinant, inhibitory anti-idiotypic antibody in monovalent Fab format; it is recommended as capture antibody in a PK bridging ELISA format with HCA329P for detection.
- Product Type
- Monoclonal Antibody
- Clone
- AbD34433
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA330 | E | 0.1 mg |
|
Human Anti-Ipilimumab Antibody, clone AbD34433 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the monoclonal antibody ipilimumab. It does not recognize free human CTLA-4 (cytotoxic T-lymphocyte antigen-4) or ipilimumab in complex with CTLA-4. The antibody can be used to measure free ipilimumab in patient samples.
A pair of anti-ipilimumab 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 HRP conjugated Anti-Ipilimumab Antibody in full immunoglobulin format, clone AbD34429ia (HCA329P) as the detection antibody.
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
A pair of anti-ipilimumab 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 HRP conjugated Anti-Ipilimumab Antibody in full immunoglobulin format, clone AbD34429ia (HCA329P) as the detection antibody.
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 (kappa 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 liquid.
- Preparation
- StrepTactin affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Ipilimumab
- Affinity
- The monovalent intrinsic affinity of AbD34433 was measured as KD = 0.3 nM by real time, label free molecular interaction analysis on immobilized ipilimumab.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- 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. - Guarantee
- 12 months from date of despatch
More Information
- Acknowledgements
- Sold under license of U.S. Patents 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad.
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 AbD34433 can be used in an indirect ELISA or as a capture antibody in a sandwich ELISA together with HCA329P as the detection reagent.
Protocol: PK bridging ELISA
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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 (Drug/Target Complex) | HCA331 | E | 0.1 mg |
|
|
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody |
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