Anti-GLP-1 (GLP-1(7-36)amide, C-terminal specific)

Cat.No. HYB 147-06

Cat. No.
HYB 147-06
Product name
Anti-GLP-1 (GLP-1(7-36)amide, C-terminal specific)
Description

Mouse monoclonal antibody

Specificity

HYB 147-06 is specific for the amidated C-terminus of the peptide and does not react with GLP-1(7-37) (1).  

Immunogen

Synthetic GLP-1(7-36)amide coupled to carrier 

Application
Species reactivity (positive)
Human, mouse
Species reactivity (negative)
Not determined
Subclass

IgG1/k

Gene ID
2641
Clone number
8G9
Epitope

C-terminal epitope of GLP-1(7-36)amide

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

Glucagon-like peptide 1(7-36)amide (GLP-1(7-36)amide) is the principal active form of GLP-1, the other being GLP-1(7-37). GLP-1 is a peptide hormone of the glucagon family, produced by the L cells of the intestinal mucosa from the same prohormone as glucagon. The active forms are potent stimulators of glucose-dependent insulin secretion. The sequence of GLP-1 is fully conserved in all mammalian species examined so far.

Application 1

ELISA

HYB 147-06 reacts with the amidated C-terminus of GLP-1(7-36)amide, GLP-1(9-36)amide and GLP-1(1-36)amide. HYB 147-06 can be used as capture antibody in sandwich ELISA (1) using HYB 147-12B (total GLP-1) or ABS 033-10B (active GLP-1) as detection antibody (2, 3).  
In a sandwich ELISA ABS 044-49 (as capture antibody) forms a pair with  HYB 147-06B (as biotinylated detection antibody) in order to measure ”degraded GLP-1” (the GLP-1 metabolite (GLP-1(9-36)amide)).
This uses HYB 147-06 as capture antibody in combination with biotinylated detection antibody HYB 147-12 to measure the sum of GLP-1(7-36)amide, GLP-1(1-36)amide and GLP-1(9-36)amide in biological samples. Relative to measuring the two active forms of GLP-1, the responses are reduced by not measuring the contribution of GLP-1(7- 37) to the overall response, but this is compensated for by measuring an approximately equal contribution from GLP-1(1-36)amide. However, the GLP-1(9-36)amide degradation product is also measured, augmenting the response above that of the active forms alone. This assay cannot detect changes in active GLP-1 in response to treatment with DPP-4 inhibitors, and is therefore only suitable for experiments in which changes in GLP-1 degradation are irrelevant.
This uses HYB 147-06 as capture antibody in combination with biotinylated detection antibody ABS 033-10 exclusively to measure GLP-1(7-36)amide in biological samples. GLP-1(7-36)amide is the principal active form of the peptide, and has been estimated to make up about 70% of the physiological response. Relative to measuring both active forms of GLP-1 together, the responses are reduced by not measuring the approximately 30% contribution of GLP-1(7-37) to the overall response. There is no cross-reactivity with any extended or truncated physiological form of GLP-1(7-36)amide. Specimens must be collected into tubes containing an adequate amount of DPP-4 inhibitor (in addition to routine peptidase inhibitors) to prevent the rapid in vitro degradation of the active peptide.
The calibration curve of a sandwich assay for measuring degraded GLP-1 using ABS 044-49 as the capture antibody and HYB 147-06B as the biotinylated detection antibody.
Application 2

IHC

HYB 147-06 has been used for the immunoblockade of endogenous GLP-1 in rats (2).
 
Pancreas sections from diabetic mice stained with HYB 147-06 (1:500). Courtesy of Dr. Annette Plesner, University of British Columbia

1. Ghiglione M, Uttenthal LO, Koch C (1993) Monoclonal antibodies to glucagon-like peptide-1. Digestion 54:396-397.

2. Marchetti P, Lupi R, Bugliani M, Kirkpatrick CL, Sebastiani G, Grieco FA, Del Guerra S, D'Aleo V, Piro S, Marselli L, Boggi U, Filipponi F, Tinti L, Salvini L, Wollheim CB, Purrello F, Dotta F (2012) A local glucagon-like peptide 1 (GLP-1) system in human pancreatic islets. Diabetologia 55:3262-3272.
 

3. Piotrowski K, Becker M, Zugwurst J, Biller-Friedmann I, Spoettl G, Greif M, Leber AW, Becker A, Laubender RP, Lebherz C, Goeke B, Marx N, Parhofer KG, Lehrke M (2013) Circulating concentrations of GLP-1 are associated with coronary atherosclerosis in humans. Cardiovascular Diabetology 12:117.

4.  Voortman T, Hendriks HFJ, Witkamp RF, Wortelboer HM (2012) Effects of long- and short-chain fatty acids on the relase of gastrointestinal hormones using an ex vivo porcine intestinal tissue model. J. Agric. Food Chem. 60:9035-9042.