Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Flow cytometry [1]
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Validation data
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- Product number
- 376-2239-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD223 (LAG-3) Monoclonal Antibody (3DS223H), Brilliant Ultra Violet™ 661, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This 3DS223H monoclonal antibody recognizes human CD223 also known as Lymphocyte Activation Gene 3 (LAG-3). LAG-3 is a 70-kDa surface glycoprotein belonging to the Ig superfamily with homology to CD4. LAG-3 binds to MHC class II with higher affinity than CD4 and is thought to be involved in the negative regulation of T cell activation and homeostatic proliferation. Surface expression of LAG-3 has been reported on activated T cells (including regulatory T cells) and NK cells. CD8+ T cells usually express LAG-3 at significantly higher levels than CD4+ T cells. Coexpression of LAG-3 and CD49b has been proposed to identify human and mouse Type 1 regulatory T cells (Tr1 cells). This 3DS223H antibody will recognize a formaldehyde-fixed epitope. Applications Reported: This 3DS223H antibody has been reported for use in flow cytometric analysis. Applications Tested: This 3DS223H antibody has been pre-diluted and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This may be used at 5 µL (0.25 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. Brilliant Ultra Violet™ 661 (BUV661) is a tandem dye that emits at 660 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information. 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 # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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: 350 nm; Emission: 660 nm; Laser: Ultraviolet Laser. BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 3DS223H
- 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
- Normal human peripheral blood cells were stimulated for 72 hours with CD3 Monoclonal Antibody, Functional Grade (Product # 16-0037-85), CD28 Monoclonal Antibody, Functional Grade (Product # 16-0289-85), and Human IL-2 Recombinant Protein. Cells were then stained with CD8a Monoclonal Antibody, FITC (Product # 11-0088-42) and Mouse IgG1 kappa Isotype Control, Brilliant Ultra Violet 661 (Product # 376-4714-81) (left) or CD223 (LAG-3) Monoclonal Antibody, Brilliant Ultra Violet 661 (right). Viable cells in the lymphocyte gate were used for analysis, as determined by 7-AAD (Product # 00-6993-50).