Antibody data
- Antibody Data
- Antigen structure
- References [3]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [4]
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Validation data
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- Product number
- 47-2239-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD223 (LAG-3) Monoclonal Antibody (3DS223H), APC-eFluor™ 780, 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-titrated and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.125 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. APC-eFluor® 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. 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: 633-647 nm; Emission: 780 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
- 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 MAVS Genetic Variation Is Associated with Decreased HIV-1 Replication In Vitro and Reduced CD4(+) T Cell Infection in HIV-1-Infected Individuals.
OMIP-037: 16-color panel to measure inhibitory receptor signatures from multiple human immune cell subsets.
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
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)
OMIP-037: 16-color panel to measure inhibitory receptor signatures from multiple human immune cell subsets.
Belkina AC, Snyder-Cappione JE
Cytometry. Part A : the journal of the International Society for Analytical Cytology 2017 Feb;91(2):175-179
Cytometry. Part A : the journal of the International Society for Analytical Cytology 2017 Feb;91(2):175-179
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
Miles B, Miller SM, Folkvord JM, Kimball A, Chamanian M, Meditz AL, Arends T, McCarter MD, Levy DN, Rakasz EG, Skinner PJ, Connick E
Nature communications 2015 Oct 20;6:8608
Nature communications 2015 Oct 20;6:8608
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of 3-day Anti-CD3 (Product # 16-0037-81) and Anti-CD28 stimulated (Product # 16-0289-81) normal human peripheral blood cells with Anti-Human CD8a PE (Product # 12-0086-42) and Mouse IgG1 K Isotype Control APC-eFluor® 780 (Product # 47-4714-82) (left) or Anti-Human CD223 (LAG-3) APC-eFluor® 780 (right). Viable cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 6 T FR exhibit an enhanced regulatory phenotype in ex vivo HIV infection. Tonsil cells were mock-, X4-, or R5-spinoculated and cultured under experimental conditions as indicated. T FR were then analysed for expression of regulatory receptors and cytokine production by intracellular cytokine staining. ( a ) Percentage of total (surface and intracellular) T FR CTLA-4 expression ( n =15). ( b ) Percentage of surface T FR LAG-3 expression ( n =8). ( c ) Production of IL-10 by T FR ( n =7). ( d ) Production of TGF-beta-1 (measured as LAP) by T FR ( n =5). The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Friedman ( a , b ) or Mann-Whitney tests ( c , d ) and significance is denoted by asterisks where * P
- 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.