Anti-GLP-1(7-37) and GLP-1(7-36)amide (N-terminal specific)

Cat.No. ABS 033-10

Cat. No.
ABS 033-10
Product name
Anti-GLP-1(7-37) and GLP-1(7-36)amide (N-terminal specific)
Description

Mouse monoclonal antibody

Specificity

ABS 033-10 binds the free N-terminus of GLP-1(7-37) and GLP-1(7-36)amide and shows <0.2% cross-reactivity with GLP-1(9-36)amide, glucagon and human GIP. Cross-reacts 0.25% with GLP-1(1-37), approximately 6% with human GLP-2 and approximately 50% with exendin-4.

Immunogen

Synthetic GLP-1(7-17) 

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

IgG2a/k

Gene ID
2641
Clone number
10
Epitope

Free N-terminus of GLP-1(7-37) and 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
ABS 033-10 binds free GLP-1(7-37) and free GLP-1(7-36)amide in solution.
ABS 033-10 reacts in ELISA with GLP-1(7-37) coated directly onto the microtiter well.
 
ABS 033-10 binds free exendin-4 in solution and can function as a capture antibody for exendin-4 in combination with ABS 012-35B in order to measure Exendin-4. 
 
When measureing “active” GLP-1 ABS 033-10B works as a detection antibody in combination with HYB 147-12.
 
ABS 033-10B can be used as a detection antibody in combination with ABS 046-03 as a capture antibody in order to measure non-amidated GLP-1 forms and cross-reacting about 5% with C-terminally amidated GLP-1. Results show detection limits of 44pmol/L which is 10-20 times higher than the basal concentration of GLP-1, so the assays have to be optimized. 
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 therapid in vitro degradation of the active peptide.
By using the above monoclonal antibodies, it is possible to measure intact exendin-4 by a sandwich ELISA which is not subject to interference from GLP-1 or other glucagon-related peptides that may be present. ABS 033-10 is used as capture antibody in combination with a biotinylated detection antibody from the ABS 012 series. The best sensitivity in an unoptimized buffer assay was obtained with biotinylated ABS 012-35 as the detection antibody.
The calibration curve of a sandwich assay for Non-amidated GLP-1 using ABS 046-03 as the capture antibody and ABS 033-10B as the biotinylated detection antibody.

 

Application 2

WB
ABS 033-10 can be used in Western blotting.