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AB315061

Human LAG-3 ELISA Kit

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Human LAG-3 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human LAG-3 with a sensitivity of 22.37 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Validated on a number of sample types including cerebrospinal fluid (CSF)

View Alternative Names

CD223, FDC, LAG3, Lymphocyte activation gene 3 protein, LAG-3

4 Images
Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)
  • sELISA

Supplier Data

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)

Interpolated concentration of native LAG-3 was measured in duplicate at different sample concentrations. Undiluted samples for stimulated PBMC supernatant are 100%. PBMC’s were grown in the presence of 1.5% PHA-M for 46 hours. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)
  • sELISA

Supplier Data

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)

Example of human LAG-3 standard curve. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)
  • sELISA

Supplier Data

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)

Serum of ten individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean LAG-3 concentration was determined to be 3,092 pg/mL with a range of 1,685 – 4,171 pg/mL.

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)
  • sELISA

Supplier Data

Sandwich ELISA - Human LAG-3 ELISA Kit (AB315061)

Interpolated concentration of native LAG-3 was measured in duplicate at different sample concentrations. Undiluted samples are as follows : serum 25%, plasma (citrate) 25%, plasma (EDTA) 50%, and plasma (heparin) 50%. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2).

Key facts

Detection method

Colorimetric

Sample types

Serum, Citrate plasma, Heparin Plasma, EDTA Plasma, Cell culture supernatant, Cerebral Spinal Fluid

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 22.37 pg/mL

Range

93.75 - 6000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

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Product details

Human LAG-3 SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of LAG-3 protein in Serum, Cit Plasma, Hep Plasma, EDTA Plasma, Cell culture supernatant, Cerebral Spinal Fluid. Quantitate Human LAG-3 with 22.37 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

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Precision

[ { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "5.1" }, { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.88" } ]
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Recovery

[ { "sample": "Serum", "range": "83 - 92 %", "average": "= 87" }, { "sample": "Citrate plasma", "range": "87 - 90 %", "average": "= 88" }, { "sample": "Heparin Plasma", "range": "91 - 102 %", "average": "= 98" }, { "sample": "EDTA Plasma", "range": "83 - 89 %", "average": "= 86" }, { "sample": "Cell culture supernatant", "range": "94 - 102 %", "average": "= 98" }, { "sample": "Cerebral Spinal Fluid", "range": "92 - 109 %", "average": "= 100" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

LAG-3 also known as CD223 is a transmembrane protein with a mass of approximately 70 kDa. It acts as a checkpoint molecule similar to CTLA-4 and PD-1 in the immune system. LAG-3 is expressed on activated T cells natural killer cells and some dendritic cells. It interacts with its ligand MHC class II leading to inhibition of cellular proliferation and cytokine secretion an important part of immune response regulation.
Biological function summary

LAG-3 is involved in maintaining immune homeostasis and preventing autoimmunity. It is not part of a complex but works in synergy with other immune checkpoint molecules. When interacting with MHC class II LAG-3 transmits inhibitory signals that attenuate immune cell responses. Its role in immune regulation helps to balance immune activation and tolerance which is essential for preventing tissue damage while protecting against pathogens.

Pathways

The inhibitory role of LAG-3 integrates into the T cell receptor (TCR) signaling pathway and is also linked to the PD-1 signaling pathway. In TCR signaling LAG-3 negatively regulates T cell activation. LAG-3 is functionally related to proteins like PD-1 and CTLA-4 sharing similar roles in immune checkpoint pathways that are critical for maintaining immune balance.

LAG-3 has links to cancer and autoimmune diseases. In cancer LAG-3 expression on tumor-infiltrating lymphocytes correlates with immune evasion by tumors. This relationship appears in various cancers including melanoma and renal cell carcinoma. In autoimmune diseases LAG-3's expression on regulatory T cells helps control conditions such as multiple sclerosis. It collaborates with CTLA-4 in modulating immune responses that prevent excessive immune activation and tissue damage associated with autoimmunity.
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Product protocols

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Target data

Lymphocyte activation gene 3 protein : Inhibitory receptor on antigen activated T-cells (PubMed : 20421648, PubMed : 7805750, PubMed : 8647185). Delivers inhibitory signals upon binding to ligands, such as FGL1 (By similarity). FGL1 constitutes a major ligand of LAG3 and is responsible for LAG3 T-cell inhibitory function (By similarity). Following TCR engagement, LAG3 associates with CD3-TCR in the immunological synapse and directly inhibits T-cell activation (By similarity). May inhibit antigen-specific T-cell activation in synergy with PDCD1/PD-1, possibly by acting as a coreceptor for PDCD1/PD-1 (By similarity). Negatively regulates the proliferation, activation, effector function and homeostasis of both CD8(+) and CD4(+) T-cells (PubMed : 20421648, PubMed : 7805750, PubMed : 8647185). Also mediates immune tolerance : constitutively expressed on a subset of regulatory T-cells (Tregs) and contributes to their suppressive function (By similarity). Also acts as a negative regulator of plasmacytoid dendritic cell (pDCs) activation (By similarity). Binds MHC class II (MHC-II); the precise role of MHC-II-binding is however unclear (PubMed : 8647185).. Secreted lymphocyte activation gene 3 protein. May function as a ligand for MHC class II (MHC-II) on antigen-presenting cells (APC), promoting APC activation/maturation and driving Th1 immune response.
See full target information LAG3
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websiteProtocolBooklet
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