LEGEND MAX™ Flex-T™ Human Class I Peptide Exchange ELISA Kit

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Other Names
MHC tetramer, HLA tetramer, HLA monomer, MHC monomer, biotinylated HLA monomer, Peptide exchange, Flexible peptide loading
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Flex-T™ HLA-A*02:01 UVX monomer (Cat. No. 280003) was treated with a 366nm UV in the presence of a Positive or a Negative control peptide, or test peptides (n=8), or no peptide (UV-only). After peptide exchange, each reaction mixture was then diluted following the instructions in “Reagent and Sample Preparation” section and analyzed by the ELISA. The optical densities of each sample calculated by absorbance at 450 subtracted with absorbance at 570 are shown in upper bar graph and the percentile (%) of test peptide against Positive control peptide in lower bar graph. Please refer to the BioLegend Flex-T website for details regarding data interpretation on UV-activated peptide exchange (https://www.biolegend.com/en-us/flex-t).
  • Flex-T™ HLA-A*02:01 UVX monomer (Cat. No. 280003) was treated with a 366nm UV in the presence of a Positive or a Negative control peptide, or test peptides (n=8), or no peptide (UV-only). After peptide exchange, each reaction mixture was then diluted following the instructions in “Reagent and Sample Preparation” section and analyzed by the ELISA. The optical densities of each sample calculated by absorbance at 450 subtracted with absorbance at 570 are shown in upper bar graph and the percentile (%) of test peptide against Positive control peptide in lower bar graph. Please refer to the BioLegend Flex-T website for details regarding data interpretation on UV-activated peptide exchange (https://www.biolegend.com/en-us/flex-t).
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447207 1 Pre-coated Plate 316€
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Description

BioLegend’s Flex-T™ MHC products allow high flexibility in studying antigen specific T cells. Biotinylated MHC monomers are manufactured with a UV-labile peptide that can be degraded with an ultraviolet light source. This allows for a peptide exchange when UV irradiation is performed in the presence of a peptide of interest (shown in below graph). This flexibility permits the screening of virtually any peptide of interest with enough affinity for the MHC allele that it is loaded onto. Refer to the BioLegend Flex-T website for more details (www.biolegend.com/en-us/flex-t).

BioLegend’s LEGEND MAX™ Flex-T™ HLA Class I ELISA kit is a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with a 96-well strip plate that is pre-coated with a mouse monoclonal anti-human β2-microglobulin antibody. HRP-conjugated streptavidin is the detection reagent that binds to biotinylated HLA class I monomer. Through binding of β2-microglobulin subunit and biotin conjugated on the HLA class I monomer, this Sandwich assay detects the HLA class I α-chain/β2-microglobulin/peptide complex and only intact complex is recognized. Therefore, exchange product with high affinity peptides will show relatively high signals in this assay, while moderate or low affinity peptides exchange would result in relatively lower or non-detectable signals. A Flex-T™ HLA Class I ELISA Control is included in the kit as the assay control. This kit is not intended to be used to measure the actual affinity of peptides, but is specifically designed to qualitatively evaluate the efficiency of UV peptide exchange when loading peptides of interest into Flex-T™ HLA class I monomers. Each assay kit comes with analytically validated, ready-to-use reagents.

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents
  • Flex-T™ Pre-coated 96-well Strip Microplate
  • Flex-T™ HLA Class I ELISA Control
  • Avidin-HRP
  • Assay Buffer A
  • Wash Buffer (20X)
  • Substrate Solution F
  • Stop Solution
  • Plate Sealers

Product Details

Reactivity
Human
Application

ELISA

Materials Not Included
  • Microplate reader able to measure absorbance at 450 nm
  • Adjustable pipettes to measure volumes ranging from 1 μL to 1,000 μL
  • Deionized water
  • Wash bottle or automated microplate washer
  • Polypropylene vials to prepare control dilutions
  • Timer
  • Plate Shaker

Antigen Details

Cell Type
T cells
Biology Area
Adaptive Immunity
Molecular Family
MHC Antigens
Gene ID
NA
UniProt
View information about Class I Peptide on UniProt.org

Related FAQs

For some of your ELISA kits, why do my serum samples require dilution with assay buffer?

Dilution with assay buffer is required to minimize the matrix difference between the samples and the standards to achieve better accuracy.

I have multiple LEGEND MAX™ ELISA kits that I want to run simultaneously. Can I use the same wash buffer for all the kits?

The Wash buffer is the same for all the current LEGEND MAX™ kits. All the part numbers on the Wash Buffer bottles in these kits should be the same. For ELISA MAX™ Deluxe and ELISA MAX™ Standard sets, we provide a recipe for the wash buffer on each kit’s technical data sheet. This recipe is the same for all ELISA MAX™ sets.

In your LEGEND MAX™ ELISA Kits, there is a step that calls for a washing of the plates before even adding any sample to it. What is the purpose of this step?

We typically use a stabilizer for pre-coated plates. The washings were designed to remove these components before you start the assay. If you do not do the washings, the effect on assay performance is negligible.

Go To Top Version: 1    Revision Date: 01.20.2020

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

BioLegend Inc., 8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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