Antibody data
- Antibody Data
- Antigen structure
- References [10]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [5]
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- Product number
- 46-2239-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD223 (LAG-3) Monoclonal Antibody (3DS223H), PerCP-eFluor™ 710, 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 (0.06 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. 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
- 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 RASA2 ablation in T cells boosts antigen sensitivity and long-term function.
CD4(+) LAG-3(+) T cells are decreased in active psoriatic arthritis patients and their restoration in vitro is mediated by TNF inhibitors.
Senolytic CAR T cells reverse senescence-associated pathologies.
MAVS Genetic Variation Is Associated with Decreased HIV-1 Replication In Vitro and Reduced CD4(+) T Cell Infection in HIV-1-Infected Individuals.
Effect of analytical treatment interruption and reinitiation of antiretroviral therapy on HIV reservoirs and immunologic parameters in infected individuals.
Targeted reconstruction of T cell receptor sequence from single cell RNA-seq links CDR3 length to T cell differentiation state.
Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection.
OMIP-037: 16-color panel to measure inhibitory receptor signatures from multiple human immune cell subsets.
Immunological biomarkers predict HIV-1 viral rebound after treatment interruption.
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
Carnevale J, Shifrut E, Kale N, Nyberg WA, Blaeschke F, Chen YY, Li Z, Bapat SP, Diolaiti ME, O'Leary P, Vedova S, Belk J, Daniel B, Roth TL, Bachl S, Anido AA, Prinzing B, Ibañez-Vega J, Lange S, Haydar D, Luetke-Eversloh M, Born-Bony M, Hegde B, Kogan S, Feuchtinger T, Okada H, Satpathy AT, Shannon K, Gottschalk S, Eyquem J, Krenciute G, Ashworth A, Marson A
Nature 2022 Sep;609(7925):174-182
Nature 2022 Sep;609(7925):174-182
CD4(+) LAG-3(+) T cells are decreased in active psoriatic arthritis patients and their restoration in vitro is mediated by TNF inhibitors.
Gertel S, Polachek A, Furer V, Levartovsky D, Elkayam O
Clinical and experimental immunology 2021 Nov;206(2):173-183
Clinical and experimental immunology 2021 Nov;206(2):173-183
Senolytic CAR T cells reverse senescence-associated pathologies.
Amor C, Feucht J, Leibold J, Ho YJ, Zhu C, Alonso-Curbelo D, Mansilla-Soto J, Boyer JA, Li X, Giavridis T, Kulick A, Houlihan S, Peerschke E, Friedman SL, Ponomarev V, Piersigilli A, Sadelain M, Lowe SW
Nature 2020 Jul;583(7814):127-132
Nature 2020 Jul;583(7814):127-132
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)
Effect of analytical treatment interruption and reinitiation of antiretroviral therapy on HIV reservoirs and immunologic parameters in infected individuals.
Clarridge KE, Blazkova J, Einkauf K, Petrone M, Refsland EW, Justement JS, Shi V, Huiting ED, Seamon CA, Lee GQ, Yu XG, Moir S, Sneller MC, Lichterfeld M, Chun TW
PLoS pathogens 2018 Jan;14(1):e1006792
PLoS pathogens 2018 Jan;14(1):e1006792
Targeted reconstruction of T cell receptor sequence from single cell RNA-seq links CDR3 length to T cell differentiation state.
Afik S, Yates KB, Bi K, Darko S, Godec J, Gerdemann U, Swadling L, Douek DC, Klenerman P, Barnes EJ, Sharpe AH, Haining WN, Yosef N
Nucleic acids research 2017 Sep 19;45(16):e148
Nucleic acids research 2017 Sep 19;45(16):e148
Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection.
Eyquem J, Mansilla-Soto J, Giavridis T, van der Stegen SJ, Hamieh M, Cunanan KM, Odak A, Gönen M, Sadelain M
Nature 2017 Mar 2;543(7643):113-117
Nature 2017 Mar 2;543(7643):113-117
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
Immunological biomarkers predict HIV-1 viral rebound after treatment interruption.
Hurst J, Hoffmann M, Pace M, Williams JP, Thornhill J, Hamlyn E, Meyerowitz J, Willberg C, Koelsch KK, Robinson N, Brown H, Fisher M, Kinloch S, Cooper DA, Schechter M, Tambussi G, Fidler S, Babiker A, Weber J, Kelleher AD, Phillips RE, Frater J
Nature communications 2015 Oct 9;6:8495
Nature communications 2015 Oct 9;6:8495
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
- Normal human peripheral blood cells were stimulated for 3 days with Human IL-2 Recombinant Protein (Product # 14-8029-81), Anti-Human CD3, and Anti-Human CD28 Functional Grade Purifieds (Product # 16-0037-81 and Product # 16-0289-81). These cells were then surface stained with Anti-Human CD8a APC (Product # 17-0087-42) and Mouse IgG1 K Isotype Control PerCP-eFluor® 710 (Product # 46-4714-82) (left) or Anti-Human CD223 (LAG-3) PerCP-eFluor® 710 (right). Cells in the lymphocyte gate 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
- 2 FIGURE The effect of in-vitro incubation on the %CD4 + lymphocyte-activation gene (LAG)-3 + T cell population. Peripheral blood mononuclear cells (PBMCs) of healthy controls ( n = 5) were analyzed by flow cytometry when fresh after undergoing 5 days in culture in vitro to determine the percentage of %CD4 + LAG-3 + T cells. (a,b) Representative flow cytometric dot-plot of a healthy donor's PBMCs' gating strategy presented as a side-scatter area (SSC) and a forward-scatter area (FSC) (left panels). Evaluation of %CD4 + LAG-3 + T cells (right panels) is shown. The percentage of positive cells is indicated in the upper right quadrant. (c) Graph indicating the average percentage of %CD4 + LAG-3 + T cells for tested donors ( n = 5). Data are shown with standard error of the mean (SEM) values; * p < 0.01 (Mann-Whitney U -test)
- 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.