HTRF® Anti-immunoglobulin Reagents
HTRF® toolbox reagents include secondary murine, sheep, rabbit, and human antibodies, as well as immunoglobulin binding proteins. These reagents are useful in developing new assays in which directly labeled assay components are not available or which involve chimeric proteins. Most of our anti-immunoglobulin reagents are available conjugated to either Europium-cryptate or XL665 for maximum flexibility.
- High affinity polyclonal antibodies
- Resistant to most buffer conditions and additives (e.g. DMSO, pH, chelators, ionic strength)
- Compatible with membrane and cell-based assays
- Lyophilized packaging for easy handling and long term storage
- Proven batch-to-batch reproducibility
Product Specifications:
Anti-Ig HTRF® reagents have been strictly selected based on their specificity. Polyclonal antibodies (PAb) to murine, rabbit, and human immunoglobulins (IgGs) are absorbed against most human and animal serum proteins to minimize inter-species cross-reactivity.
| Motif / Speciesn | Antibody | Species | Specificity |
| Mouse immunoglobulins | PAb | Rabbit | Mouse Igs |
| Rabbit immunoglobulins | PAb | Goat | Rabbit Igs |
| Human immunoglobulins | PAb | Goat | Human Fc Igs |
Assay Performance:
CD28 provides a major costimulatory signal for CD4-positive T cells. Ligation with its natural ligands CD80 (B7-1) and CD86 (B7-2) leads to signals during activation that are required for the production of interleukin-2. This process has been implicated in the regulation of T-cell anergy and programmed cell death. The HTRF anti-immunoglobulin reagents were used to develop an assay for antagonists of CD28 and CD86 binding. Both molecules were respectively tagged with human and rat IgG Fc fragments and detected using the XL665-labled anti-human antibody. CD86 was expressed as a fusion protein with a rat Ig domain which is recognized by a biotinylated sheep anti-rat antibody followed by binding of europium-labeled streptavidin. Results of the binding assay were published by Mellor, et.al.
Figure 1: Data for the CD28-CD86 binding assay is published in: Mellor et al. Development of a CD28/CD86 (B7-2) binding assay for high throughput screening by homogenous time-resolved fluorescence. J Biomol Screening. 1998; 3: 91-99.
Ordering Info:
Reagents are sold by the number of tests (384-well, 20 µl reactions). Antibodies conjugated to XL665 are supplied on the basis of 20ng of antibody per well. The active moiety per vial is also provided. In practice, the active moiety amount is generally preferred to that of total conjugate as a basis for calculating assay development.
For more information about using HTRF toolbox reagents, click here.
Anti-immunoglobulin Reagents:
Europium Conjugates:
| Description | Quantity* | Active Moiety/vial | Cat no. | Product Insert |
| Anti-mouse Ig K | 5,000 tests | 7.5 µg | 61PAMKLA | |
| Anti-mouse Ig K | 20,000 tests | 30 µg | 61PAMKLB | |
| Anti-rabbit Ig K | 5,000 tests | 7.5 µg | 61PARKLA! | |
| Anti-rabbit Ig K | 20,000 tests | 30 µg | 61PARKLB | |
| Anti-human Ig K | 5,000 tests | 10 µg | 61HFCKLA | — |
| Anti-human Ig K | 20,000 tests | 40 µg | 61HFCKLB | — |
| Protein A K | 5,000 tests | 3.5 µg | 61PRAKLA ** | — |
| Protein A K | 20,000 tests | 14 µg | 61PRAKLB ** | — |
XL665 Conjugates:
| Description | Quantity* | Active Moiety/vial | Cat no. | — |
| Anti-mouse Ig XL665 | 5,000 tests | 100 µg | 61PAMXLA | |
| Anti-mouse Ig XL665 | 20,000 tests | 400µg | 61PAMXLB | |
| Anti-rabbit Ig XL665 | 5,000 tests | 100 µg | 61PARXLA | — |
| Anti-rabbit Ig XL665 | 20,000 tests | 400 µg | 61PARXLB | — |
| Anti-human Ig XL665 | 5,000 tests | 100 µg | 61HFCXLA | — |
| Anti-human Ig XL665 | 20,000 tests | 400 µg | 61HFCXLB | — |
| Protein A XL665 | 5,000 tests | 50 µg | 61PRAXLA * | — |
| Protein A XL665 | 20,000 tests | 200 µg | 61PRAXLB * | — |
| *based on 20 µl assay volume **upon request |
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For more information or To Place an Order:
U.S. and Canada
1-888-963-4567
Europe and other countries
+33(0)466 796 705
Japan
+81-3-5510-2932
