Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Flow cytometry [2]
- Other assay [2]
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Validation data
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- Product number
- 64-2239-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD223 (LAG-3) Monoclonal Antibody (3DS223H), Super Bright™ 645, 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). 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 (0.5 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. Super Bright 645 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 645 nm. We recommend using a 660/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining 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) (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 degrees 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: 405 nm; Emission: 645 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 3DS223H
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Longitudinal single-cell profiling reveals molecular heterogeneity and tumor-immune evolution in refractory mantle cell lymphoma.
MAVS Genetic Variation Is Associated with Decreased HIV-1 Replication In Vitro and Reduced CD4(+) T Cell Infection in HIV-1-Infected Individuals.
Zhang S, Jiang VC, Han G, Hao D, Lian J, Liu Y, Zhang R, McIntosh J, Wang R, Dang M, Dai E, Wang Y, Santos D, Badillo M, Leeming A, Chen Z, Hartig K, Bigcal J, Zhou J, Kanagal-Shamanna R, Ok CY, Lee H, Steiner RE, Zhang J, Song X, Nair R, Ahmed S, Rodriquez A, Thirumurthi S, Jain P, Wagner-Bartak N, Hill H, Nomie K, Flowers C, Futreal A, Wang L, Wang M
Nature communications 2021 May 17;12(1):2877
Nature communications 2021 May 17;12(1):2877
MAVS Genetic Variation Is Associated with Decreased HIV-1 Replication In Vitro and Reduced CD4(+) T Cell Infection in HIV-1-Infected Individuals.
Stunnenberg M, van Pul L, Sprokholt JK, van Dort KA, Gringhuis SI, Geijtenbeek TBH, Kootstra NA
Viruses 2020 Jul 16;12(7)
Viruses 2020 Jul 16;12(7)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells were stimulated with CD3 and CD28 Monoclonal Antibodies, Functional Grade (Product # 16-0037 and 16-0289) and then stained with CD8 Monoclonal Antibody, PE (Product # 12-0086) and Mouse IgG1 kappa Isotype Control, Super Bright 645 (Product # 64-4714) (left) or CD223 Monoclonal Antibody, Super Bright 645 (right). Cells in the lymphocyte gate were used for analysis.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells were stimulated with CD3 and CD28 Monoclonal Antibodies, Functional Grade (Product # 16-0037-81 and Product # 16-0289-81) and then stained with CD8 Monoclonal Antibody, PE (Product # 12-0086-42) and Mouse IgG1 kappa Isotype Control, Super Bright 645 (Product # 64-4714-82) (left) or CD223 Monoclonal Antibody, Super Bright 645 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 MAVS genetic variation does not affect T cell exhaustion, activation, and senescence. ( a ) Percentages of PD-1 + , LAG-3 + , CD38 + , HLA-DR + , CD134 + , exhausted (PD-1 + LAG-3 + ), activated (CD38 + HLA-DR + ), and senescent (CD27 - CD28 - ) cells within CD4 + and ( b ) CD8 + T cells of untreated HIV-1-infected individuals with a MAVS minor or MAVS major genotype 2.5-3.5 years p.SC were analyzed using flow cytometry. Each square or dot represents a different study participant (median +- IQR). No significant differences between HIV-1-infected individuals with a MAVS minor or MAVS major genotype were observed.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4 MAVS minor genotype is associated with a decreased percentage of naive CD4 + T cells. ( a ) Percentages and ( b ) cell counts of naive (T naive ; CD45RA + CD27 + CCR7 + ), terminally differentiated effector memory (TEMRA; CD45RA + CCR7 - CD27 - ), central memory (CM; CD45RA - CCR7 + CD27 + ), transitional memory (TM; CD45RA - CCR7 - CD27 + ), and effector memory (EM; CD45RA - CCR7 - CD27 - ) cells within CD4 + and CD8 + T cells were analyzed using flow cytometry. ( c ) Percentages of exhausted (PD-1 + LAG-3 + ) and activated (CD38 + HLA-DR + ) CD4 + T cells and ( d ) CD8 + T cells within T naive , TEMRA, CM, TM, and EM populations were analyzed using flow cytometry. Each square or dot represents a different study participant (median +- IQR). All significant differences are indicated: * p < 0.05, unpaired Mann-Whitney test.