LAG-3/CD223 Polyclonal antibody

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  WB analysis using 16616-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 16616-1-AP (LAG-3/CD223 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse liver using 16616-1-AP

  IP result of anti-LAG-3 (IP:16616-1-AP, 4ug; Detection:16616-1-AP 1:500) with mouse liver tissue lysate 4000ug.

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  IHC staining of human tonsillitis using 16616-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 16616-1-AP (LAG-3/CD223 antibody) at dilution of 1:2000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human tonsillitis using 16616-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 16616-1-AP (LAG-3/CD223 antibody) at dilution of 1:2000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of human tonsillitis using 16616-1-AP

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using LAG-3 antibody (16616-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of human tonsillitis using 16616-1-AP

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using LAG-3 antibody (16616-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

LAG-3/CD223 Polyclonal Antibody for WB, IHC, IF-P, IP, ELISA

Cat No. 16616-1-AP

Publications(14)

Reviews (2)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF-P, IP, ELISA

Synonyms

CD223, LAG3, FDC, LAG 3, LAG-3

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Tested Applications

• Positive WB detected in: PHA treated human PBMCs
• Positive IP detected in: mouse liver tissue
• Positive IHC detected in: human tonsillitis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human tonsillitis tissue

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 7 publications below
• IHC: See 6 publications below
• IF: See 3 publications below

Product Information

16616-1-AP targets LAG-3/CD223 in WB, IHC, IF-P, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag9909

  Product name: Recombinant human LAG3 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 27-360 aa of BC052589

  Sequence: AEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITGQPQVGKE

• Full Name: lymphocyte-activation gene 3
• Calculated Molecular Weight: 525 aa, 57 kDa
• Observed Molecular Weight: 60-70 kDa
• GenBank Accession Number: BC052589
• Gene Symbol: LAG-3
• Gene ID (NCBI): 3902
• RRID: AB_2133350
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P18627
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

LAG-3, also known as CD223, is an immune checkpoint molecule that regulates both T-cell activation and homeostasis. LAG-3 is expressed on activated T cells, NK cells, regulatory T cells, and plasmacytoid dendritic cells. It is a CD4-related molecule that binds MHC class II. LAG-3 plays an important role in modulating T cell expansion and function, and blockade of LAG-3 with monoclonal antibodies can augment T cell function. (PMID: 15634887; 21086108; 28783703)

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB	Cancer Immunol Immunother; Novel roles of VAT1 expression in the immunosuppressive action of diffuse gliomas.; Authors - Pei Yang
human	IHC	Ann Transl Med; Single-cell RNA-seq reveals transcriptional landscape and intratumor heterogenicity in gallbladder cancer liver metastasis microenvironment.; Authors - Yigang Zhang
human	IHC	Front Oncol; The immune checkpoint expression in the tumor immune microenvironment of DLBCL: Clinicopathologic features and prognosis; Authors - Jiajia Ma
human	WB	Int J Oncol; Effect of DPP4/CD26 expression on SARS‑CoV‑2 susceptibility, immune response, adenosine (derivatives m62A and CD) regulations on patients with cancer and healthy individuals; Authors - Jiaman Du
human	WB	Br J Ophthalmol; Implications of LAG3 and CTLA4 immune checkpoints beyond PD-1/PD-L1 as a potential target in determining the prognosis of uveal melanoma patients; Authors - Seema Kashyap
human	WB	J Clin Invest; Targeting fibrinogen-like protein 1 enhances immunotherapy in hepatocellular carcinoma; Authors - Mingen Lin

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Sofia (Verified Customer) (05-27-2025)

Antibody did not work as expected even using the suggested antigen retrieval. I'll try it again using a different antigen retrieval.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:200
• Cell Tissue Type: Brain samples

FH

Eman (Verified Customer) (09-20-2021)

the antibody works very well for WB the recommended dilution is 1:500 to 1:1000 i think we can even dilute it more this antibody is higly recommended

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000 overnight
• Cell Tissue Type: HEK293T cells

LAG-3/CD223 Antibody Western Blot validation (1:1000 overnight dilution) in HEK293T cells (Cat no:16616-1-AP)