IgG1 antibody | AbD27686
A microtiter plate was coated overnight with various antibodies at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Human IgG1 (Fc) Antibody (clone AbD27686, HCA285) titrated to the given concentrations, and Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent (BUF049A), and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with various antibodies at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Human IgG1 (Fc):HRP Antibody (clone AbD27686, HCA285P) titrated to the given concentrations in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with VEGF at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, 10% cynomolgus macaque serum was added spiked with increasing concentrations of bevacizumab (hIgG1/kappa). Detection was performed using either HRP conjugated Human Anti-Human hIgG1 (Fc) (clone AbD27686, HCA285P), HRP conjugated Mouse Anti-Human IgG (CH2 Domain) Antibody (clone R10Z8E9, MCA5748G) or, as a control for an antibody that is not human Fc specific, Mouse Anti-Human IgG (Fc) CH2 Domain:HRP (clone MK 1 A6, MCA647P) 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 Mouse Anti-Human IgG (CH2 domain) Antibody (clone R10Z8E9, MCA5748G) at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of hIgG1 kappa antibody (0.5 μg/ml) plus increasing concentrations of either Human Anti-Human IgG1 (Fc) Antibody (clone AbD27686) or antibody clone R10Z8E9 was added. Free hIgG1 kappa antibody, still capable of binding to the R10Z8E9 coated plate, was detected using HRP conjugated R10Z8E9 antibody 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 IgG1/k, IgG1/l, cyno IgG, IgG2/k or IgG4/k antibodies at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a dilution series of biotin conjugated Human Anti-Human IgG1 (Fc) antibody (clone AbD27686, conjugated to biotin via the SpyTag/SpyCatcher protein ligation, HCA285B) in HISPEC Assay Diluent (BUF049A) was added. Detection was performed using HRP conjugated NeutrAvidin (1:8,000 in HISPEC Assay Diluent (BUF049A)) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated over night with Human Recombinant VEGF (PHP293) at a concentration of 5 µg/ml. After washing and blocking with PBST+ 5% BSA, 10% cynomolgus monkey serum was added, spiked with increasing concentrations of bevacizumab (hIgG1/kappa). Primary detection antibodies were added: either biotinylated Human Anti-Human IgG1 (Fc) Antibody (clone AbD27686, conjugated to biotin via the SpyTag/SpyCatcher protein ligation, HCA285B), or biotin conjugated Mouse Anti-Human IgG (CH2 Domain) Antibody (clone R10Z8E9) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) for 1 h at RT. Detection was performed using HRP conjugated NeutrAvidin (1:8,000 in HISPEC Assay Diluent (BUF049A)) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
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E ResetHuman anti Human IgG1 (Fc):HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD27686
- Isotype
- HuCAL Fab monovalent
- Specificity
- IgG1
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| Human Anti-Human IgG1 Fc Antibody clone AbD27686 is a recombinant monovalent Fab antibody, that recognizes human IgG1 Fc, with an affinity of 76 nM. It has been shown to have a greatly reduced affinity (89-fold) for IgG from cynomolgus macaque compared to human IgG1. It also has strongly reduced binding to rhesus macaque IgG. It does not recognize other human immunoglobulin isotypes (IgG2, IgG3, IgG4, IgA, IgE, IgM) or mouse IgG2a. This antibody has been generated by Bio-Rad using the HuCAL® antibody library and CysDisplay® technology. It is sold for research use, with implicit permission for use for the detection of therapeutic antibodies from samples obtained from experimental animals. When tested by Bio-Rad in an antigen capture pharmacokinetic assay format, this antibody performs similarly to Mouse Anti-Human IgG (CH2 Domain) Antibody clone R10Z8E9 (MCA5748G) in 10% cynomolgus macaque serum. It recognizes a different epitope on human IgG1 Fc than clone R10Z8E9. View a summary of anti-human IgG Fc specific antibodies for the detection of therapeutic antibodies. |
- Target Species
- Human
- Product Form
- A monovalent human recombinant Fab (lambda light chain) selected from the HuCAL PLATINUM phage display library. Expressed in E. coli and purified using purified using NiNTA affinity chromatography. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. Conjugated to horseradish peroxidase (HRP) - liquid.
- Buffer Solution
- Phosphate buffered saline.
- Preservative Stabilisers
- 0.01% Thiomersal.
- Immunogen
- Recombinant human IgG1 kappa
- Affinity
- The intrinsic affinity of the monovalent form of this antibody is KD=76 nM as measured by real time, label free molecular interaction analysis on immobilized hIgG1 kappa.
- Approx. Protein Concentrations
- Antibody concentration 0.1 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.
This product is shipped frozen.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
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 |  |
1/100 | 1/1000 |
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.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
| List Price | Your Price | ||||||
| Log in | |||||||
| Description | Hispec Assay Diluent | ||||||
- Licensed Use
- For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
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