Anti-GLP-1 (Mid-molecule specific)

Cat.No. HYB 147-12

Cat. No.
HYB 147-12
Product name
Anti-GLP-1 (Mid-molecule specific)
Description

Mouse monoclonal antibody

Specificity

Reacts with all forms of GLP-1, including precursor and GLP-1(9-37) /GLP-1(9-36amide) metabolite HYB 147-12 cross-reacts with Liraglutide. 

Immunogen

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

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

IgG1/k

Gene ID
2641
Clone number
4F3
Epitope

Mid-molecular epitope of GLP-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

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 147-12 binds to GLP-1 when coated directly onto the microtiter well, and binds GLP-1(7-36)amide in solution giving a Ka of 4.0x10E8 in inhibition ELISA. HYB 147-12 cross-reacts <0.4% with coated glucagon.  
In inhibition ELISA no binding of free glucagon in solution is detected, giving an estimated cross-reactivity of <0.2%. 
Biotinylated HYB 147-12 is the preferred detection antibody for measuring C-terminally amidated forms of GLP-1 in combination with HYB 147-06 as capture antibody (1). HYB 147-12 can be used as a capture antibody in combination with ABS 046-03B as a detection antibody for measuring non-amidated GLP-1 forms and cross- reacting about 16% 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 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.
Application 2
WB
In Western blotting a dilution guideline of 1/2000 has proved successful.
HYB 147-12 is applicable for immunoaffinity.
Application 3

IHC

Although not tested, HYB 147-12 is likely to detect all known molecular forms of GLP-1 in immunohistochemistry. 

1. 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.