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MABF954 Anti-LAG3 Antibody, clone 4-10-C9

This Anti-LAG3 Antibody, clone 4-10-C9 is validated for use in Flow Cytometry, Immunocytochemistry for the detection of LAG3.

Anti-LAG3 Antibody, clone 4-10-C9 MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Inflammation & Immunology Research Sub-Category: Immunological Signaling Gene Symbol: Lag3 UniProt Number: Q61790
MABF954
100 μg  
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Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
MFC, ICCMPurifiedMonoclonal Antibody
Description
Catalogue NumberMABF954
DescriptionAnti-LAG3 Antibody, clone 4-10-C9
Alternate Names
  • Lymphocyte activation gene 3 protein
  • CD223
  • LAG-3
Background InformationLymphocyte 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).
References
Product Information
FormatPurified
PresentationPurified mouse monoclonal IgG2aκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationThis Anti-LAG3 Antibody, clone 4-10-C9 is validated for use in Flow Cytometry, Immunocytochemistry for the detection of LAG3.
Key Applications
  • Flow Cytometry
  • Immunocytochemistry
Application NotesFlow 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).
Biological Information
ImmunogenMurine LAG-3-expressing mouse T-cell hybridoma.
EpitopeWithin D3/D4 domains.
Clone4-10-C9
ConcentrationPlease refer to lot specific datasheet.
HostMouse
SpecificityClone 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).
IsotypeIgG2aκ
Species Reactivity
  • Mouse
Species Reactivity NoteMouse.
Antibody TypeMonoclonal Antibody
Entrez Gene Number
Gene Symbol
  • Lag3
Purification MethodProtein G purified.
UniProt Number
Molecular Weight54.51/56.98 kDa (mature/pro-form) calculated.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceEvaluated 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.
Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
Packaging Information
Material Size100 μg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
MABF954 04055977352924

Documentation

Anti-LAG3 Antibody, clone 4-10-C9 SDS

Title

Safety Data Sheet (SDS) 

Anti-LAG3 Antibody, clone 4-10-C9 Certificates of Analysis

TitleLot Number
Anti-LAG3, clone 4-10-C9 - 3384837 3384837
Anti-LAG3, clone 4-10-C9 - 3703580 3703580
Anti-LAG3, clone 4-10-C9 - 3794318 3794318
Anti-LAG3, clone 4-10-C9 - 3884758 3884758
Anti-LAG3, clone 4-10-C9 -Q2690232 Q2690232
Anti-LAG3, clone 4-10-C9 Monoclonal Antibody 2943580
Anti-LAG3, clone 4-10-C9 Monoclonal Antibody 3009979
Anti-LAG3, clone 4-10-C9 Monoclonal Antibody 2874600
Anti-LAG3, clone 4-10-C9 Monoclonal Antibody 2930964
Anti-LAG3, clone 4-10-C9 Monoclonal Antibody 2902696