Recombinant human LAG3 (Lymphocyte activation gene 3 protein, also known as CD223), encompassing amino acids 23-450. The C-terminal portion of this construct is fused with the Fc fragment of a human IgG1 followed by an Avi-Tag™. The recombinant protein was enzymatically biotinylated using the Avi-Tag™ and affinity purified. HiP™ indicates a high purity protein (≥90% pure) and less than 10% aggregation as measured by gel filtration.
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Synonyms
Lymphocyte-Activation Gene 3, CD223, Lag, Lag-3
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Product Data Gallery
4-20% SDS-PAGE Coomassie Staining
LAG3-FGL1 Activity
Biotin-Avidin Pulldown
Gel Filtration Trace
FGL1:LAG3 inhibition curve using a TR-FRET assay (#79739-2). Representative data only; data may vary by lot.
20 mM NaOAC, pH 5.5, 500 mM NaCl, 0.02% Tween 20 and 10% glycerol
MW
75 kDa + glycans
Amino Acids
23-450
Glycosylation
This protein runs at a higher MW by SDS-PAGE due to glycosylation.
Aggregation
<10%
Genbank #
NM_002286
UniProt #
P18627
Tag(s)
C-terminal Avi-Tag
Label
This protein is enzymatically biotinylated using Avi-Tag™ technology. Biotinylation is confirmed to be ≥90%. For more information on enzymatic biotinylation, please see our Tech Note.
References
1. Byrnes, et al., Cytometry A. 2012 Sep;81(2):165-8. 2. Di Carlo, et al., J. Pathol. 2005 Jan; 205(1):82-91.
Storage/Stability
At least 6 months at –80°C.
Assay Conditions
Assay was conducted in 1x Immuno Buffer 1 (BPS Bioscience #79311). 5 µl FGL1 His from Cloud Clone Corp. #RPD022Hu01 P20211110840 was used with a final concentration of 100 nM along with 5 µl of titrated LAG3 (CD223), Biotin-labeled (Human) HiP™ (BPS Bioscience #71147) titrated from 150-0 ng/rxn and incubated at room temperature for 1 hour. 10 µl 1:250-fold diluted Nickel Chelate acceptor beads Perkin Elmer #AL108C were added to the plate and incubated at room temperature for 30 min. 10 µl 1:125 fold diluted Strep Donor Beads Perkin Elmer #6760002S were added and incubated at room temperature for 1 hour. Α-counts were read using a Tecan infinite M1000 Pro plate reader.
Instructions for Use
Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.
Applications
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.