Anti-Gliadin peptide

Cat.No. HYB 314-01

Cat. No.
HYB 314-01
Product name
Anti-Gliadin peptide
Description

Mouse monoclonal antibody

Specificity

HYB 314-01 reacts strongly with the deamidated peptide but cross-reacts about 5% with the non-deamidated peptide KLQPFPQPQLPYPQPQ.

Immunogen

Gliadin-related peptide Lys57-Glu65-[alpha-gliadin (58-73)] (KLQPFPQPELPYPQPQ) 

Application
Species reactivity (positive)
Wheat, Rye, Barley
Species reactivity (negative)
Oat
Subclass

IgG2a/k

Clone number
14D5
Epitope

The epitope is located in a central region of the Lys57-Glu65-[alpha-gliadin (58-73)] (KLQPFPQPELPYPQPQ) peptide (1).

Content

Available in 200 µL and 1 mL size.1 mg/mL +/- 15%. See Certificate of Analysis for details.

Purification
Protein-A purified
Form
Liquid
Solvent
0.01 M phosphate buffer, pH 7.4, containing 0.5 M NaCl and 15 mM sodium azide
Storage
4-8ºC without exposure to light. No precautions necessary during handling.
Target

Celiac disease is associated with a CD4+ T-cell response to epitopes of gliadin presented by HLA-DQ2 or -DQ8 class II MHC molecules. These epitopes are present in a 33-mer peptide of wheat alpha-gliadin, residues 56-88, which is resistant to digestion and forms a substrate for tissue transglutaminase (TG2), generating the glutamic acid residues essential for binding to HLA-DQ2. The immunogen corresponds to a deamidated form of a region that includes the T-cell epitopes, including the immunodominant PQPQLPY region and two PXPQP motifs associated with binding to IgA from patients with celiac disease. Complete homology exists between residues 63-73, 70-80 and 77-87 of wheat alpha gliadin.

Application 1

ELISA
HYB 314-01 reacts with both the deamidated peptide (KLQPFPQPELPYPQPQ) and non-deamidated petide (KLQPFPQPQLPYPQPQ) when these are coated directly in an immunoplate, but approx. 20-fold higher concentrations of antibody are needed to produce the same signal with the non-deamidated peptide. (2)

Application 2

WB
HYB 314-01 can be used in western blotting.
 

A western blot of Gliadin peptide (HYB 314-01) measured in rye (lane 1), barley (lane 2) and wheat (lane 3). The SDS gel was run under reducing conditions, and HYB 314-01 diluted 1:1000.

1. Hartwig Petersen N, Hansen PR, Houen G (2011) Fast and efficient characterization of an anti-gliadin monoclonal antibody epitope related to celiac disease using resin-bound peptides. J immunol Methods 365:174-182.

2. Skovbjerg H, Koch C, Anthonsen D, Sjostrom H (2004) Deamidation and cross-linking of gliadin peptides by transglutaminases and the relation to celiac disease. Biochim Biophys Acta 1690:220-230.

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