"I have used a wide variety of secondaries and Jackson ImmunoResearch has consistently been the best. The fluorophores are bright and stable and their selective (x reactivity removed) secondaries have always shown species specificity in multiple labeling."
Janet Duerr, Ohio UniversityRating: 5.0
Jackson ImmunoResearch Mouse Monoclonal Anti-Human IgE antibodies are the latest addition to our suite of products suitable for diagnostics research and development. Specific for Human IgE antibodies, they complement our existing Anti-Human IgG, IgM, and IgA antibodies. Anti-Human IgE is available conjugated to a select range of reporter molecules, including Alkaline Phosphatase and Biotin enabling excellent sensitivity.
Jackson ImmunoResearch Anti-Human IgE is a mouse-derived monoclonal antibody with reactivity to human E class immunoglobulins and is epsilon (ε) chain specific.
Jackson ImmunoResearch Mouse Anti-Human IgE (clone number ME.114) (209-002-241) is available conjugated to a range of reporter molecules and is suitable to detect human IgE by a variety of techniques, including ELISA (Enzyme-linked immunosorbent assay), Lateral flow immunoassay (LFIA), flow cytometry, western blot, and CLIA (Chemiluminescent immunoassay).
Jackson ImmunoResearch Anti-Human IgE epsilon (ε) chain specific and has been tested by ELISA and shows minimal cross-reactivity to human IgG, IgM, or IgA. Figure 1 is a Western blot that demonstrates the specificity of Jackson ImmunoResearch Anti-Human IgE showing no detection of other immunoglobulin classes. We have not tested if this antibody will detect immunoglobulins from other species.
Jackson ImmunoResearch Anti-Human IgE can be used to detect human IgE from various sources with excellent sensitivity. Figure 3 demonstrates the sensitivity of Jackson ImmunoResearch Anti-Human IgE when used in an indirect ELISA, showing its ability to detect IgE coating the wells at two different concentrations. Figure 4 demonstrates the sensitivity of Jackson ImmunoResearch Anti-Human IgE when used in an indirect ELISA to detect IgE purified from different myeloma and plasma sources.
Jackson ImmunoResearch Anti-Human IgE offers excellent sensitivity when used in a sandwich ELISA with its capture partner antibody. We recommend using two different monoclonal antibodies for capture and detection. In this format, use an unconjugated Anti-Human IgE as your coating antibody before blocking with Bovine Serum Albumin (IgG-Free, Protease-Free 001-000-161). Following washing, incubate with your diluted sample. For the detection step, use a conjugated Anti-Human IgE antibody, such as Alkaline Phosphatase Anti-Human IgE (209-052-241) or Peroxidase Anti-Human IgE (209-032-241), followed by the appropriate substrate or visualization technique. Figure 5 illustrates the application of the direct sandwich assay to validate the quantification of total IgE from patient samples using unconjugated Anti-Human IgE in partnership with an alkaline phosphatase conjugated detection antibody. Total IgE (kIU/l) in patient samples used in this experiment were previously measured by the Roche Cobas Total IgE assay. These results are displayed on the x-axis of Figure 5.
The dilution factors suggested in the following table are presented as ranges because the optimal dilution is a function of many factors, such as antigen density, permeability, etc. The optimal working dilution should be determined empirically for each application.
| Application | |||
|---|---|---|---|
| Conjugate | ELISA | Western Blots* | Flow Cytometry |
| Unconjugated | 10-20 µg/ml | ||
| FITC | 1:50-1:200 | ||
| R-PE | 1:50-1:200 | ||
| Horseradish Peroxidase | 1:5,000-1:100,000 | 1:5,000-1:100,000 (non-ECL) 1:10,000-1:200,000 (ECL) |
|
| Alkaline Phosphatase | 1:5,000-1:50,000 | 1:5,000-1:50,000 | |
| Biotin-SP (using Fluorophore-Conjugated Streptavidin) | 1:200-1:1,000 | ||
| Biotin-SP (using enzyme-Conjugated Streptavidin) | 1:20,000-1:400,000 | 1:20,000-1:400,000 | |
* Please note that this antibody will give a weaker signal when detecting reduced protein and therefore is not recommended for applications where detection of reduced IgE is required by western blot.
