产品详情
产品参数(Product Specifications)
Assay TypeSandwich-ELISAAnalyteMHC class I Monomer (HLA-A*02:01) folded with peptidesFormat96testsReactivityHumanRegulatory StatusRUOStandard Curve Range0.048-200 μMAssay Time6 hrSuitable Sample TypePeptides that can bind to MHC class I monomer (HLA-A*02:01)Sample volume100 μL产品概述(Product Overview)
The screening of peptides constitutes a critical step in the development of vaccines and therapeutic approaches such as TCR-T therapy. High-throughput screening kits facilitate the rapid and convenient selection of target peptides.
应用说明(Application)
The kit is developed for screening MHC binding peptides, and evaluated the peptide affinity of HLA-A*02:01 alleles.
It is for research use only.
存储(Storage)
1. Store the unopened kit at 2-8°C upon receipt.
2. Locate the expiration date on the outer packaging and do not use reagents beyond their expiration date.
3. The opened kit should be stored according to the storage conditions listed in the Components Table. The shelf life of the opened kit is 30 days from the date of opening.
组分(Materials Provided)
IDComponentsSizeHEP-0201-C01Pre-Coated Anti-MHC class I Antibody Microplate1 plate (8×12 strips)HEP-0201-C02Biotinylated MHC class I Monomer (HLA-A*02:01)120 μgHEP-0201-C03HRP Conjugated Detection Antibody12 μgHEP-0201-C04HLA-A*02:01 Reference Peptide1 mgHEP-0201-C05Peptide Exchange Factor10 mLHEP-0201-C06Assay Buffer50 mLHEP-0201-C0720×Washing Buffer50 mLHEP-0201-C08Substrate Solution12 mLHEP-0201-C09Stop Solution8 mL原理(Assay Principles)
The kit contains two modules:
1) To generate a monomer with new peptide specificity, the biotinylated MHC class I monomer (HLA-A*02:01), which is initially folded with a low-affinity existing peptide, is incubated with a peptide of interest in the presence of a proprietary Peptide Exchange Factor.
2) A simple colorimetric sandwich ELISA assay containing a Pre-Coated Anti-MHC class I Antibody Microplate to capture MHC class I Monomers and a HRP Conjugated Detection Antibody recognizing the exiting peptide. This assay allows to measure the ability of test peptides to bind HLA-A*02:01 by quantifying the displacement of the exiting peptide. Your experiment will include 3 simple steps:
a) Add your new generated MHC monomers loaded with specific peptides of interest to the pre-coated microplate.
b) Add HRP Conjugated Detection Antibody.
c) Add TMB or other colorimetric HRP substrate. The absorbance value is negatively correlated with the efficiency of peptide exchange. amount of the specific peptides of interest exchanged onto the MHC monomer.
When the exiting peptide is replaced by the peptide of interest, the HRP Conjugated Detection Antibody can’t couple to the plate so that the absorbance value is low. On the contrary, high absorbance indicates low peptide exchange.

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数据展示
典型数据-Typical Data
Please refer to DS document for the assay protocol.

Reference peptide exchange with Biotinylated MHC class I Monomer (HLA-A*02:01)
Mix the serial dilutions of HLA-A*02:01 Reference Peptide (HEP-0201-C04) (1:3 serial dilution, from 200μM to 0.05μM) with Biotinylated MHC class I Monomer (HLA-A*02:01) (HEP-0201-C02). The IC50 of the sandwich ELISA assay is 6.237μM. Typically, the Peptide Exchange Rate of 200μM is 93.1%. (QC tested).
A high concordance rate of 88% was observed between the predicted and measured IC50 values for the tested peptides (37 out of 42).
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