Mouse / Rat GLP-1 Active (7-36) ELISA Kit

$580.00

The Eagle Biosciences Mouse/Rat Active GLP-1 ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of bioactive glucagon-like peptide-1 (7-36) amide [GLP-1 (7-36)] level in rat and mouse plasma samples with only 20 µl of sample volume. Eagle Biosciences Mouse / Rat Active GLP-1 ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Mouse/Rat GLP-1 Active (7-36) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.1 pM
Dynamic Range: 0.85 – 146 pM
Incubation Time: Overnight
Sample Type: Plasma*  
Sample Size: 20 µL

Controls Included

GLP-1(7-36) pg/ml = GLP-1 (7-36) pM x 3.298
*The primary amino acid sequence of GLP-1 peptide is identical among mammalian species, i.e. rat, mouse, pig, human, etc.

Product Developed and Manufactured in the USA


Additional Information

Assay Principle

The Eagle Biosciences Mouse / Rat Active GLP-1 ELISA Assay Kit is designed, developed and produced for the quantitative measurement of bioactive GLP-1 (7-36) in rodent plasma where there is usually have limited amount of sample available for analysis.  The assay utilizes the two-site “sandwich” technique with two selected GLP-1 (7-36) specific antibodies.

Assay standards, controls and test samples are directly added to wells of a microplate that is coated with streptavidin. Subsequently, a mixture of biotinylated GLP-1 (7-36) specific antibody and a horseradish peroxidase (HRP) conjugated GLP-1 (7-36) specific antibody is added to each well. After the first incubation period, a “sandwich” immunocomplex of “Streptavidin – Biotin-Antibody – GLP-1(7-36) – HRP conjugated antibody” is formed and attached to the wall of the plate. The unbound HRP conjugated antibody is removed in a subsequent washing step. For the detection of this immunocomplex, each well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to GLP-1 (7-36) on the wall of the microtiter well is directly proportional to the amount of GLP-1 (7-36) in the sample.

Place a sufficient number of streptavidin coated microwell strips/wells in a holder to run GLP-1 (7-36) standards, controls and unknown samples in duplicate.

Assay Procedure

  1. Prepare GLP-1 (7-36) Antibody Mixture: mixing GLP-1 Tracer Antibody and Capture Antibody by 1:21 fold dilution of the Tracer Antibody and by 1:21 fold dilution of the biotinylated Capture Antibody with the Tracer antibody Diluent. For each strip, it is required to mix 1 mL of the Tracer Antibody Diluent with 50 µL the Capture Antibody and 50 µL of the Tracer Antibody in a clean test tube.
  2. Add 20 µL of standards, controls and test samples into the designated microwell.
  3. Add 100 µL of GLP-1 (7-36) Antibody Mixture to each well
  4. Cover the plate with one plate sealer and incubate plate at 2-8 °C, static for 20 – 24 hours.
  5. Remove plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  6. Add 100 µL of ELISA HRP Substrate into each of the wells.
  7. Cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
  8. Incubate plate at room temperature, static for 20 min.
  9. Remove the aluminum foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently
  10. Read the absorbance at wavelength 450 nm/620 nm or 450 nm/650 nm within 10 minutes in a microplate reader

Typical Standard Curve

GP121-K01 GLP-1 ELISA-Mouse

Manual

Product Manual