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LAG-3 Recombinant Rabbit Monoclonal Antibody [PSH0-44]

Usd: 385

Specification

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Catalog# HA721346

LAG-3 Recombinant Rabbit Monoclonal Antibody [PSH0-44]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

LAG-3 Recombinant Rabbit Monoclonal Antibody [PSH0-44]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human LAG-3 aa 1-500 / 525.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 57 kDa

Positive Control

NIH/3T3 transfected with LAG3 cell lysate, human tonsil tissue, NIH/3T3 transfected with LAG3.

Conjugation

unconjugated

Clone Number

PSH0-44

RRID

AB_3072463

Reactivity Data

WB IHC-P IF-Cell
Human
Not Recommended
Tested Tested
Mouse
Tested Tested
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200

  • IF-Cell

  • 1:100

Target

Function

Lymphocyte-activation gene 3, also known as LAG-3, is a protein which in humans is encoded by the LAG3 gene. LAG3, which was discovered in 1990 and was designated CD223 (cluster of differentiation 223) after the Seventh Human Leucocyte Differentiation Antigen Workshop in 2000, is a cell surface molecule with diverse biologic effects on T cell function. It is an immune checkpoint receptor and as such is the target of various drug development programs by pharmaceutical companies seeking to develop new treatments for cancer and autoimmune disorders. In soluble form it is also being developed as a cancer drug in its own right. LAG3's main ligand is MHC class II, to which it binds with higher affinity than CD4. The protein negatively regulates cellular proliferation, activation, and homeostasis of T cells, in a similar fashion to CTLA-4 and PD-1 and has been reported to play a role in Treg suppressive function. Fibrinogen-like protein1 FGL1, a liver-secreted protein, is another (major) LAG3 functional ligand independent of MHC-II. LAG3 also helps maintain CD8+ T cells in a tolerogenic state and, working with PD-1, helps maintain CD8 exhaustion during chronic viral infection. LAG3 is known to be involved in the maturation and activation of dendritic cells.

Background References

1. Chocarro L et al. Understanding LAG-3 Signaling. Int J Mol Sci. 2021 May

2. Maruhashi T et al. LAG-3: from molecular functions to clinical applications. J Immunother Cancer. 2020 Sep

Subcellular Location

Cell membrane; Secreted.

UNIPROT

SwissProt: P18627 Human

SwissProt: Q61790 Mouse

Synonyms

CD223 antibody

CD223 antigen antibody

FDC protein antibody

LAG-3 antibody

Lag3 antibody

LAG3_HUMAN antibody

Lymphocyte activating 3 antibody

Lymphocyte activation gene 3 antibody

Lymphocyte activation gene 3 protein antibody

Protein FDC antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of LAG-3 on different lysates with Rabbit anti-LAG-3 antibody (<a href="/products/HA721346" style="font-weight: bold;text-decoration: underline;">HA721346</a>) at 1/2,000 dilution.<br /><br />Lane 1: NIH/3T3 transfected with LAG3 expression vector, whole cell lysate<br />Lane 2: NIH/3T3 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 57 kDa<br />Observed band size: 55/70 kDa<br /><br />Exposure time: 1 minute 10 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721346" style="font-weight: bold;text-decoration: underline;">HA721346</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of LAG-3 on different lysates with Rabbit anti-LAG-3 antibody (HA721346) at 1/2,000 dilution.

    Lane 1: NIH/3T3 transfected with LAG3 expression vector, whole cell lysate
    Lane 2: NIH/3T3 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 57 kDa
    Observed band size: 55/70 kDa

    Exposure time: 1 minute 10 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721346) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LAG-3 antibody (<a href="/products/HA721346" style="font-weight: bold;text-decoration: underline;">HA721346</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721346" style="font-weight: bold;text-decoration: underline;">HA721346</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LAG-3 antibody (HA721346) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721346) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of NIH/3T3 cells transfected with or without LAG3 labeling LAG-3 with Rabbit anti-LAG-3 antibody (<a href="/products/HA721346" style="font-weight: bold;text-decoration: underline;">HA721346</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LAG-3 antibody (<a href="/products/HA721346" style="font-weight: bold;text-decoration: underline;">HA721346</a>) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of NIH/3T3 cells transfected with or without LAG3 labeling LAG-3 with Rabbit anti-LAG-3 antibody (HA721346) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-LAG-3 antibody (HA721346) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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