PROVETOP

Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer Protein

Catalog No: HLAG-HP005

Species
Human
Expression System
HEK293
Tag
His, Avi
Activity
Activity verified

Product overview

Recombinant Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer Protein is expressed from HEK293 with His tag and Avi tag at the C-terminus,tetramer is assembled by biotinylated monomer and streptavidin. It contains Gly25-Thr305(HLA-G), Ile21-Met119(B2M) and RIIPRHLQL peptide (accession: P17693-1(HLA-G) & P61769(B2M) &RIIPRHLQL).

Product Details

Molecular Aliases
HLA G antigen; sHLA-G; b2 microglobulin; HLA G; HLAG; HLA-G; MHC Class I Antigen G; MHC class Ib antigen; MHC-G; sHLA-G
Protein Length
Gly25-Thr305
Expression System
HEK293
Theoretical Molecular Weight
The protein has a predicted MW of 258 kDa. Due to glycosylation, the protein migrates to 260-265 kDa under Non reduced (N) condition based on Bis-Tris PAGE result.
Purity
> 95% as determined by Bis-Tris PAGE > 90% as determined by HPLC
Endotoxin
Less than 1 EU per μg by the LAL method.
Buffer / Formulation
Lyophilized from 0.22μm filtered solution in PBS, 100mM L-Arginine (pH 7.4). Normally 8% trehalose is added as protectant before lyophilization.
State
Lyophilized
Storage Conditions
-20 to -80°C for 12 months as supplied from date of receipt. -80°C for 3 months after reconstitution. Recommend to aliquot the protein into smaller quantities for optimal storage. Please minimize freeze-thaw cycles.
Reconstitution Advice
Dissolve the lyophilized protein in distilled water. Please refer to the Certificate of Analysis for detailed instructions.

Data Display

Bis-Tris PAGE
HLAG-HP005 Bis-Tris-PAGE-white result

Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer on Bis-Tris PAGE under Non reduced (N) condition. The purity is greater than 95%.

SEC-HPLC
HLAG-HP005 SEC-HPLC result

The purity of Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer is greater than 90% as determined by SEC-HPLC.

ELISA
HLAG-HP005 ELISA result

Immobilized Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer, His Tag at 0.5μg/ml (100μl/well) on the plate. Dose response curve for Anti-HLA-G Antibody, hFc Tag with the EC50 of 6.2ng/ml determined by ELISA (QC Test).

Blocking
HLAG-HP005 Blocking result

Serial dilutions of Anti-LILRB2 Antibody were added into Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer, His Tag: Human LILRB2, mFc Tag binding reactioins. The half maximal inhibitiory concentration (IC50) is 0.11μg/ml.

SPR
HLAG-HP005 SPR result

Human LILRB2, hFc Tag captured on CM5 Chip via Protein A can bind Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer with an affinity constant of 4.62 nM as determined in SPR assay (Biacore T200).

SPR
HLAG-HP005 SPR-2 result

Human HLA-G&B2M&Peptide (RIIPRHLQL) Tetramer, His Tag immobilized on CM5 Chip can bind Human LILRB2 Domain1&2, His Tag with an affinity constant of 6.5μM as determined in a SPR assay (Biacore T200).

Background

HLA-G is a molecule that was first known to confer protection to the fetus from destruction by the immune system of its mother, thus critically contributing to fetal-maternal tolerance. The first functional finding constituted the basis for HLA-G research and can be summarized as such: HLA-G, membrane-bound or soluble, strongly binds its inhibitory receptors on immune cells (NK, T, B, monocytes/dendritic cells), inhibits the functions of these effectors, and so induces immune inhibition.

References

  1. (1) Krijgsman D, Roelands J, Hendrickx W, Bedognetti D, Kuppen PJK. HLA-G: A New Immune Checkpoint in Cancer? Int J Mol Sci. 2020 Jun 25;21(12):4528. doi: 10.3390/ijms21124528. PMID: 32630545; PMCID: PMC7350262.

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