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LAG-3 Antibody [Unconjugated]

Images

 
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 µg/mL of ...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
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
LAG‑3 was detected in immersion fixed paraffin-embedded sections of human spleen using Goat Anti-Human LAG‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) at 3 µg/mL for 1 ...read more
LAG‑3 was detected in immersion fixed paraffin-embedded sections of mouse spleen using Goat Anti-Human LAG‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2319) at 15 µg/mL for 1 ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB, Flow, IHC, CyTOF-ready
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.
Source
N/A
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Gene
LAG3
Purity Statement
Antigen Affinity-purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • CyTOF-reported
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 3-15 ug/mL
  • Western Blot 1 ug/mL
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 either lyophilized or 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
  • LAG3
  • LAG-3
  • lymphocyte activating 3
  • lymphocyte activation gene 3 protein
  • lymphocyte-activation gene 3
  • Secreted lymphocyte activation gene 3 protein
  • sLAG-3

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 4 applications: Flow Cytometry, Functional Assay, ICC, Western Blot.


Filter By Application
Flow Cytometry
(3)
Functional Assay
(2)
ICC
(1)
Western Blot
(1)
All Applications
Filter By Species
Human
(5)
All Species
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., 2018-12-03;0(0):. 2018-12-03 [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, 2017-03-20;543(7647):728-732. 2017-03-20 [PMID: 28321130] (Functional Assay, Human) Functional Assay Human
Fine needle aspirate flow cytometric phenotyping characterizes immunosuppressive nature of the mesothelioma microenvironment Sci Rep, 2016-08-19;6(0):31745. 2016-08-19 [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, 2014-08-08;193(6):3101-12. 2014-08-08 [PMID: 25108024] (Functional Assay, ICC, Western Blot, Human) Functional Assay, ICC, Western Blot Human
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, 2011-06-07;118(4):965-74. 2011-06-07 [PMID: 21652684] (Flow Cytometry, Human) Flow Cytometry Human

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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for LAG-3 Antibody (AF2319). (Showing 1 - 2 of 2 FAQs).

  1. 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).
  2. 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.

Secondary Antibodies

 

Isotype Controls

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Blogs on LAG-3.

Tired T cells: Hypoxia Drives T cell Exhaustion in the Tumor Microenvironment
By Hunter MartinezThe paradigm shifting view of the immune system being leveraged to target cancer has led to numerous therapeutic breakthroughs. One major cell group responsible for this revelation is a T cell. ...  Read full blog post.

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Bioinformatics

Gene Symbol LAG3
Uniprot