Antibody data
- Antibody Data
- Antigen structure
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- Comments [0]
- Validations
- Flow cytometry [1]
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- Product number
- 46-9753-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SEMA4A Monoclonal Antibody (5E3), PerCP-eFluor™ 710, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The monoclonal antibody 5E3 reacts with human and mouse Semaphorin 4A (Sema4A). This 150 kDa glycoprotein is a member of the class IV semaphorin family and exists as membrane-bound and soluble forms. Sema4A is expressed by a wide variety of tissues, including lung, kidney, spleen, testis, and mammary glands, as well as activated T cells and dendritic cells. This ligand binds to Tim-2, neuropilin-1 (Nrp1), plexin D1, and plexin B1. Sema4A plays a role in T cell activation, Th1 differentiation, and Treg function and survival. Our testing demonstrates that the 5E3 antibody stains resting CD49b+ NK cells in mouse. However, staining of CD56+ NK cells in human is negative. Applications Reported: This 5E3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 5E3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 5E3
- Vial size
- 25 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Staining of normal human peripheral blood cells with Anti-Human CD11c PE (Product # 12-0116-42) and Mouse IgG1 K Isotype Control PerCP-eFluor® 710 (Product # 46-4714-82) (left) or Anti-Human/Mouse Semaphorin 4A PerCP-eFluor® 710 (right). Cells in the lymphocyte gate were used for analysis.