Histamine Assay
Histamine is a small biogenic amine known to play an important role in immediate hypersensitivity and inflammation reactions as well as several atopic diseases such as allergic rhinitis, bronchial asthma and eczema. Cisbio Bioassays developed a highly specific functional assay for quantification of histamine that requires only a single microplate including acylation and detection. This HTRF®-based Histamine assay is easily miniaturized and amenable to HTS.
- Homogeneous assay
- Single plate protocol
- Highly accurate
- Amenable to automation
- Histamine measurement
- Allergy & Asthma assessment
Assay Principle:
The HTRF® histamine assay is an immunocompetitive assay which uses XL665-labeled histamine and an anti-histamine Cryptate-labeled antibody. The assay has a two-step procotol: acylation and detection. Acylation and detection can both be carried out in a single plate for simplified automation.
Product Specifications:
| Detection limit: | 1.8 nM histamine |
| Dynamic range: | 10 - 400 nM histamine |
| EC50 histamine: | 43 nM |
| Specificity: | Acylated histamine 100% Acylated L-histidine < 0.00067% Acylated serotonin < 0.0067% |
Figure 1: Histamine standard curves were performed using the designated final assay volumes and plate types.
Assay Performance:
Rat basophils (RBL-2H3) were cultured in the presence or absence of 2.5 µM ionomycin to stimulate histamine release. Total histamine content was determined by lysing cells and spontaneous release was evaluated by incubating the cells in medium only. The results are shown in Figure 2.
Similar studies were performed using human peripheral blood basophils from two individual donors (figure 3). Non-specific release of histamine using Ionomycin was compared to specific degranulation that was induced by cross-linking a membrane-bound IgE with anti-IgE. Error bars represent a standard deviation of 3 replicates.
Figure 2: Histamine detection was performed following 30 minute cell stimulation at 37ºC using the standard HTRF histamine assay protocol. Emissions were read at 620 and 665nm and histamine concentration determined.
Figure 3: Histamine detection was performed using the standard HTRF histamine assay protocol following 30 minutes of stimulation of peripheral blood basophils at 37ºC. Emissions were read at 620 and 665nm and histamine concentration determined.
Ordering Info:
Histamine Assay Reagents:
| Description | Quantity* | Cat no. | Product Insert | MSDS | ||
| Histamine kit | 1,000 tests | 62HTMPEB | Request MSDS | |||
| Histamine bulk kit | 20,000 tests | 62HTMPEC | Request MSDS | |||
| *based on 20 µl assay volume | ||||||
Companion Products:
| Description | Quantity* | Cat no. |
| Histamine calibrator | 62HTMCDA | |
| Histamine control | 62HTMTDA | |
| Diluent for standard curve preparation | 62DL2DDD | |
| Acylation reagent for Histamine assay | 1,000 tests (10.5 mg/vial) |
62HTMADA |
| 20,000 tests (211 mg/vial) |
62HTMADC | |
| Acylation buffer for Histamine assay | 1,000 tests (2 mg/vial) |
62HTMFDA |
| 20,000 tests (40 mg/vial) |
62HTMFDD |
For more information or To Place an Order:
Europe and other countries
+33(0)466 796 705
