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Invitrogen
CD223 (LAG-3) Monoclonal Antibody (3DS223H), NovaFluor™ Yellow 590, eBioscience™
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FIGURE: 1 / 3
CD223 (LAG-3) Antibody (H048T03Y02-A) in Flow
3-day Anti-Human CD3 and CD28 Functional Grade Purified (Product # 16-0037-81 and Product # 16-0289-81)-stimulated normal human peripheral blood cells were unstained (left) or stained with CD223 Monoclonal Antibody, NovaFluor Yellow 590 (right). All cells were co-stained with CD8a Monoclonal Antibody, eFluor 450 (Product # 48-0086-42). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
H048T03Y02-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
3DS223H
Conjugate
Excitation/Emission Max
563/594 nm
View spectra
Form
Liquid
Concentration
0.4 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3098580 (AB_2925965)
Product Specific Information
Description
This 3DS223H monoclonal antibody recognizes human CD223 also known as Lymphocyte Activation Gene 3 (LAG-3). Coexpression of LAG-3 and CD49b has been proposed to identify human and mouse Type 1 regulatory T cells (Tr1 cells). This 3DS223H antibody will recognize a formaldehyde-fixed epitope.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This 3DS223H antibody has been pre-titrated and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 590 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 563 nm; Emission: 594 nm; Laser: 561 nm (Yellow) Laser
Target Information
LAG-3 is a 70-kDa surface glycoprotein in the immunoglobulin superfamily, with structural homology to CD4. It binds to MHC class II molecules with higher affinity than CD4 and is involved in the negative regulation of T cell activation and homeostatic proliferation. LAG-3 is expressed on the surface of activated T cells, including regulatory T cells, and NK cells. Notably, CD8+ T cells express LAG-3 at significantly higher levels than CD4+ T cells. Additionally, the coexpression of LAG-3 and CD49b has been proposed as a marker for identifying Type 1 regulatory T cells (Tr1 cells) in both humans and mice. This highlights LAG-3s role in immune regulation and its potential as a target for therapeutic interventions in immune-related conditions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: CD223; LAG-3; Lymphocyte activation gene 3 protein; lymphocyte-activation gene 3; sLAG 3; sLAG3; soluble LAG 3lymphocyte activating 3; soluble LAG3
Gene Aliases: CD223; FDC; LAG3
UniProt ID: (Human) P18627
Entrez Gene ID: (Human) 3902
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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