LAG-3 Antibody [Unconjugated]

Images

 
HEK293 human embryonic kidney cell line transfected with either (A) human LAG-3 or (B) irrelevant transfectants and eGFP was stained with Goat Anti-Human LAG-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # ...read more
Western blot shows lysates of human peripheral blood mononuclear cells (PBMC) untreated or treated (+) with 1 ug/mL PHA for 5 days and HDLM‑2 human Hodgkin's lymphoma cell line. PVDF membrane was probed with 1 ...read more
Human peripheral blood mononuclear cells (PBMCs) either (A) untreated or (B) treated with 1 ug/mL PHA for 5 days were stained with Goat Anti-Human LAG‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB, Flow, CyTOF-reported
Clonality
Polyclonal
Host
Goat
Conjugate
Unconjugated
Concentration
LYOPH

Order Details

LAG-3 Antibody [Unconjugated] Summary

Immunogen
Mouse myeloma cell line NS0-derived recombinant human LAG-3
Leu23-Leu450
Accession # P18627
Specificity
Detects human LAG-3 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse LAG-3 is observed.
Details of Functionality
ActivityAssaywCitation not found None None
Source
N/A
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Gene
LAG3
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • CyTOF-reported
Control
LAG-3 Knockout HeLa Cell Lysate
Publications
Read Publications using
AF2319 in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Preservative
No Preservative
Concentration
LYOPH
Reconstitution Instructions
Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for LAG-3 Antibody [Unconjugated]

  • CD223 antigen
  • CD223
  • FDC
  • LAG3
  • LAG-3
  • lymphocyte activation gene 3 protein
  • lymphocyte-activation gene 3
  • Protein FDC

Background

LAG-3 (Lymphocyte activation gene-3), also known as CD223, is a member of the immunoglobulin superfamily (IgSF). The mature LAG-3 protein is a 496 amino acid (aa) membrane protein with a 421 aa extracellular region which contains four IgSF domains, a 21 aa transmembrane region and a 54 aa cytoplasmic region. LAG-3 and CD4 molecules share < 20% aa sequence homology but have a similar structure (1, 2). Both molecules bind to MHC class II. LAG-3 binds to MHC class II with higher affinity compared to CD4. Both LAG-3 and CD4 genes are located on the distal part of the short arm of chromosome 12. LAG-3 is an activation-induced molecule, expressed on activated T cells and NK cells, but not on resting T cells. Studies using LAG-3 -/- mice have shown significant delay of T cell apoptosis following antigen stimulation and increased size of memory T cells pool following infection (3, 4). It also has been reported that anti-LAG-3 antibodies up-regulate T cell activation by blocking interaction of LAG-3 and MHC class II. The study has demonstrated that LAG-3 is selectively expressed on activated CD4+CD25+ TReg cells and plays a role in their suppressive activity (5). This evidence indicated, unlike the interaction of CD4 with MHC class II that plays a positive role in T cell activation, LAG-3 binds to MHC class II and negatively regulates T cell activation through LAG-3 signaling. On the other hand, studies have shown that binding of LAG-3 to MHC class II molecules on antigen presenting cells induce maturation of dendritic cells and cytokine secretion by monocytes through MHC class II signal transduction (6). Taken together, LAG-3 may have two major functions, it negatively regulates T cells activation through LAG-3 signaling and stimulates antigen presenting cells which express MHC class II.
  1. Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
  2. Baixeras, E. et al. (1992) J. Exp. Med 176:327.
  3. Workman, C.J. and D.A. Vignali (2003) Eur. J. Immunol. 33:970.
  4. Workman, C.J. et al. (2004) J. Immunol. 172:5450.
  5. Huang, C.T. et al. (2004) Immunity 21:503.
  6. Andreae, S. et al. (2003) Blood 102:2130.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for LAG-3 Antibody (AF2319)(5)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: Flow Cytometry, Functional Assay.


Filter By Application
Flow Cytometry
(3)
Functional Assay
(1)
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Filter By Species
Human
(4)
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Showing Publications 1 - 5 of 5.
Publications using AF2319 Applications Species
W Scheper, S Kelderman, LF Fanchi, C Linnemann, G Bendle, MAJ de Rooij, C Hirt, R Mezzadra, M Slagter, K Dijkstra, RJC Kluin, P Snaebjorns, K Milne, BH Nelson, H Zijlmans, G Kenter, EE Voest, JBAG Haanen, TN Schumacher Low and variable tumor reactivity of the intratumoral TCR repertoire in human cancers Nat. Med., 2018;0(0):. 2018 [PMID: 30510250] (Flow Cytometry, Human) Flow Cytometry Human
X Lu, JW Horner, E Paul, X Shang, P Troncoso, P Deng, S Jiang, Q Chang, DJ Spring, P Sharma, JA Zebala, DY Maeda, YA Wang, RA DePinho Effective combinatorial immunotherapy for castration-resistant prostate cancer Nature, 2017;543(7647):728-732. 2017 [PMID: 28321130] (Functional Assay, Human) Functional Assay Human
Fine needle aspirate flow cytometric phenotyping characterizes immunosuppressive nature of the mesothelioma microenvironment Sci Rep, 2016;6(0):31745. 2016 [PMID: 27539742] (Flow Cytometry, Human) Flow Cytometry Human
Bae J, Lee S, Park C, Lee Y, Chun T Trafficking of LAG-3 to the surface on activated T cells via its cytoplasmic domain and protein kinase C signaling. J Immunol, 0;193(6):3101-12. 0 [PMID: 25108024]
Porichis F, Kwon DS, Zupkosky J, Tighe DP, McMullen A, Brockman MA, Pavlik DF, Rodriguez-Garcia M, Pereyra F, Freeman GJ, Kavanagh DG, Kaufmann DE Responsiveness of HIV-specific CD4 T cells to PD-1 blockade. Blood, 2011;118(4):965-74. 2011 [PMID: 21652684] (Flow Cytometry, Human) Flow Cytometry Human

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for LAG-3 Antibody (AF2319). (Showing 1 - 4 of 4 FAQs).

  1. What is the concentration of your biotin conjugated LAG-3 antibody?
    • We actuall do not determine the concentration of our conjugated antibodies but they are usuall 80-100% the concentration of the base product, so the concentration should be at least 0.8 mg/ml.
  2. Can this LAG-3 antibody be used in ELISA?
    • We have not validated this particular LAG-3 antibody for ELISA in mouse; however, it has been cited as being used as a detection antibody in a publicatoin (PMID 15946792).
  3. I'm using your LAG-3 antibody in western blot but the observed band is a different size than the predicted band size, do you know what the bands are?
    • There are several alternative splice varients of LAG-3 that could explain the different sized proteins stained by the LAG-3 antibody. It is also believed that there are two secreted forms of LAG-3, one that is about 36 kDa and another at 52 kDa.
  4. Is this LAG-3 antibody offered in a smaller size that is less expensive?
    • We do not offer a sample size for this particular LAG 3 antibody but we do offer another, AF3329, in a 25 ug sample size.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

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Bioinformatics

Gene Symbol LAG3
Uniprot