HLA-A*02:01 Peptide Screening ELISA Kit

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HEP-0201-96tests
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Product Specifications

Assay Type
Sandwich-ELISA
Analyte
MHC class I Monomer (HLA-A*02:01) folded with peptides
Format
96tests
Reactivity
Human
Regulatory Status
RUO
Standard Curve Range
0.048-200 μM
Assay Time
6 h
Suitable Sample Type
Peptides that can bind to MHC class I monomer (HLA-A*02:01)
Sample volume
100 μL

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.

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.

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.

Materials Provided

ID
Components
Size
HEP-0201-C01
Pre-Coated Anti-MHC class I Antibody Microplate
1 plate (8×12 strips)
HEP-0201-C02
Biotinylated MHC class I Monomer (HLA-A*02:01)
120 μg
HEP-0201-C03
HRP Conjugated Detection Antibody
12 μg
HEP-0201-C04
HLA-A*02:01 Reference Peptide
1 mg
HEP-0201-C05
Peptide Exchange Factor
10 mL
HEP-0201-C06
Assay Buffer
50 mL
HEP-0201-C07
20×Washing Buffer
50 mL
HEP-0201-C08
Substrate Solution
12 mL
HEP-0201-C09
Stop Solution
8 mL

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.

Assay Principles