MABF954
Anti-LAG3 Antibody, clone 4-10-C9
clone 4-10-C9, from mouse
Synonym(s):
Lymphocyte activation gene 3 protein, CD223, LAG-3
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
4-10-C9, monoclonal
species reactivity
mouse
technique(s)
flow cytometry: suitable
immunocytochemistry: suitable
isotype
IgG2aκ
NCBI accession no.
UniProt accession no.
shipped in
wet ice
target post-translational modification
unmodified
Gene Information
mouse ... Lag3(16768)
General description
Lymphocyte activation gene 3 protein (UniProt Q61790; also known as CD223, LAG-3) is encoded by the Lag3 gene (Gene ID 16768) in murine species. LAG-3 is an inhibitory T-cell surface molecule that modulates T-cell activation and homeostasis. LAG-3 co-localizes with CD8 and CD4 upon TCR engagement and alters TCR signaling. LAG-3 suppresses homeostasis proliferation (HP) of both lymphocytes and some dendritic cell populations in vivo, and enhanced HP is observed in mice deficient in LAG-3. LAG-3 signaling is shown to negatively regulate STAT5 phosphorylation in activated T cells, and no enhancement of HP was seen upon LAG-3 blockade in mice deficient in both STAT5a and STAT5b. Murine LAG-3 is produced with a signal peptide sequence (a.a. 1-22), the removal of which yields the mature protein with a large extracellular region (a.a. 23-442), followed by a transmembrane segment (a.a. 443-463) and a cytoplasmic tail (a.a. 464-521). The extracellular portion contains a V-type Ig-like domain (a.a. 37-163), followed by three C2-type Ig-like domains (a.a. 165-246, 258-341, and 345-412) and elven tandem repeats of 2-amino acid E-X seqeunce at its C-terminal end (a.a. 493-518).
Specificity
Clone 4-10-C9 immunostained surface LAG-3 on activated CD4+ T cells by targeting an extracellular epitope within the third and fourth Ig-like domains (D3/D4 domains; second and third C2-type Ig-like domains). Surface LAG-3 degradation by pronase treatment abolished cell surface staining (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Immunogen
Epitope: Within D3/D4 domains.
Murine LAG-3-expressing mouse T-cell hybridoma.
Application
Flow Cytometry Analysis: 2.5 µg/mL from a representative lot detected surface LAG-3 immunoreactivity among the CD4+ and CD8+ populations of wild-type, but not Lag3-knockout, mouse splenocytes activated in vitro via CD3 cross-linking (Courtesy of Dario A. Vignali, Ph.D., University of Pittsburgh, PA, U.S.A.).
Flow Cytometry Analysis: A representative lot was fluorescently conjugated and detected an increased number of LAG-3-positive cells within the CD4+ and CD8+ populations of infiltrating lymphocytes (TILs) in tumors developed in mice exografted with murine B16 melanoma, MC38 colon adenocarcinoma, or Sa1N fibrosarcoma cells (Woo, S.R., et al. (2012). Cancer Res. 72(4): 917–927).
Flow Cytometry Analysis: A representative lot, pre-conjugated with Alexa Fluor™ 647, detected both surface and intracellular LAG-3 by immunofluorescent staining of non-permeabilized and permeabilized primary murine CD4+ T cells activated in vitro via CD3 & CD28 cross-linking by immobilized antibodies. Pronase treatment of cells prior to permeabilization abolished cell surface staining (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Flow Cytometry Analysis: A representative lot, pre-conjugated with Alexa Fluor 647, detected a time-dependent recovery of cell surface LAG-3 immunoreactivity on activated murine CD4+ T cells after initial surface LAG-3 degradation by pronase treatment. Protein synthesis inhibitor cycloheximide (Cat. No. 239764) or protein transport inhibitor Brefeldin A (Cat. No. 203729) treatment partially blocked the recovery (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Immunocytochemistry Analysis: A representative lot detected both surface and intracellular LAG-3 by fluorescent immunocytochemistry staining of non-permeabilized and permeabilized primary murine CD4+ T cells activated in vitro via CD3 & CD28 cross-linking by immobilized antibodies. Pronase treatment of cells prior to permeabilization abolished cell surface staining (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Immunocytochemistry Analysis: A representative lot detected intracellular LAG-3 immunoreactivity co-localized with those of the early and recycling endosome marker EEA1, as well as endosomal markers Rab11b and Rab27a by fluorescent immunocytochemistry staining of activated murine CD4+ T cells following pronase treatment and permeabilization (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Flow Cytometry Analysis: A representative lot was fluorescently conjugated and detected an increased number of LAG-3-positive cells within the CD4+ and CD8+ populations of infiltrating lymphocytes (TILs) in tumors developed in mice exografted with murine B16 melanoma, MC38 colon adenocarcinoma, or Sa1N fibrosarcoma cells (Woo, S.R., et al. (2012). Cancer Res. 72(4): 917–927).
Flow Cytometry Analysis: A representative lot, pre-conjugated with Alexa Fluor™ 647, detected both surface and intracellular LAG-3 by immunofluorescent staining of non-permeabilized and permeabilized primary murine CD4+ T cells activated in vitro via CD3 & CD28 cross-linking by immobilized antibodies. Pronase treatment of cells prior to permeabilization abolished cell surface staining (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Flow Cytometry Analysis: A representative lot, pre-conjugated with Alexa Fluor 647, detected a time-dependent recovery of cell surface LAG-3 immunoreactivity on activated murine CD4+ T cells after initial surface LAG-3 degradation by pronase treatment. Protein synthesis inhibitor cycloheximide (Cat. No. 239764) or protein transport inhibitor Brefeldin A (Cat. No. 203729) treatment partially blocked the recovery (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Immunocytochemistry Analysis: A representative lot detected both surface and intracellular LAG-3 by fluorescent immunocytochemistry staining of non-permeabilized and permeabilized primary murine CD4+ T cells activated in vitro via CD3 & CD28 cross-linking by immobilized antibodies. Pronase treatment of cells prior to permeabilization abolished cell surface staining (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
Immunocytochemistry Analysis: A representative lot detected intracellular LAG-3 immunoreactivity co-localized with those of the early and recycling endosome marker EEA1, as well as endosomal markers Rab11b and Rab27a by fluorescent immunocytochemistry staining of activated murine CD4+ T cells following pronase treatment and permeabilization (Woo, S.R., et al. (2010). Eur. J. Immunol. 40(6):1768-1777).
This Anti-LAG3 Antibody, clone 4-10-C9 is validated for use in Flow Cytometry, Immunocytochemistry for the detection of LAG3.
Quality
Evaluated by Flow Cytometry in mouse splenocytes.
Flow Cytometry Analysis: 1 µg/mL of this antibody detected an induction of LAG-3-positive population in isolated mouse splenocytes following a 3-day 2 µg/mL Concanavalin A (Con A) stimulation.
Flow Cytometry Analysis: 1 µg/mL of this antibody detected an induction of LAG-3-positive population in isolated mouse splenocytes following a 3-day 2 µg/mL Concanavalin A (Con A) stimulation.
Target description
54.51/56.98 kDa (mature/pro-form) calculated.
Physical form
Format: Purified
Legal Information
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Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
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Timo Eninger et al.
Proceedings of the National Academy of Sciences of the United States of America, 119(24), e2119804119-e2119804119 (2022-06-07)
Single-cell transcriptomics has revealed specific glial activation states associated with the pathogenesis of neurodegenerative diseases, such as Alzheimer’s and Parkinson’s disease. While these findings may eventually lead to new therapeutic opportunities, little is known about how these glial responses are
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