GLP-1 ELISA Kit

Catalog No. ABIN2014345

Product Details GLP-1 ELISA Kit

Target: GLP-1 (Glucagon-like peptide 1 (GLP-1))

Binding Specificity: AA 7-36, AA 9-36, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.6-54.0 pM/L

Minimum Detection Limit: 0.6 pM/L

Application: ELISA

Purpose: This ELISA (enzyme-linked immunosorbent assay) kit is produced forthe quantitative determination of the sum value of glucagon-likepeptide-1 (7-36) and (9-36) [GLP-1 (7-36)] and (9-36) [GLP-1 (9-36)] inplasma samples. It is for in vitro diagnostic use only.

Brand: EDI™

Sample Type: Plasma

Analytical Method: Quantitative

Specificity: This Total GLP-1 assay specifically measures GLP-1 (7-36) and (9-36).

Cross-Reactivity (Details): The cross reactivity of this test is listed below:GLP-1 (7-36) 100 % GLP-1 (9-36) 100 % GLP-1 (9-37) < 0.1 % GLP-1 (7-37) < 0.1 % GLP-1 (1-36) < 0.1 % GLP-2 < 0.1 %Glucagon < 0.1 %

Components: 1. Streptavidin Coated Microplate
One vial containing 12 mL ready-to-use buffer. It should be used only for tracer antibody dilution according to the assay procedures. This reagent should be stored at 2-8 °C and is stable until the expiration date on the kit box.

Material not included: 1. Precision single channel pipettes capable of delivering 25 µL, 50 µL, 100 µL, and 1000 µL etc.
2. Repeating dispenser suitable for delivering 100 µL.
3. Disposable pipette tips suitable for above volume dispensing.
4. Disposable 12 x 75 mm or 13 x 100 glass/plastic tubes.
5. Disposable plastic 100 mL and 1000 mL bottle with caps.
6. Aluminum foil.
7. Deionized or distilled water.
8. Plastic microtiter well cover or polyethylene film.
9. ELISA plate shaker.
10. ELISA multichannel wash bottle or automatic (semi-automatic) washing system.
11. Spectrophotometric microplate reader capable of reading absorbance at 450 nm.
12. DPP-4 Inhibitor.

Application Details

Sample Volume: 100 μL

Assay Time: 1 d

Plate: Pre-coated

Protocol: This ELISA is designed, developed and produced for the quantitativemeasurement of GLP-1 (7-36) and (9-36) in plasma sample. The assayutilizes the two-site sandwich technique with two selected GLP-1antibodies. This assay uses the same assay calibrators and tracerantibodies as the Active GLP-1 (7-36) ELISA (EDI™ catalog: KT-871).Assay standards, controls and test samples are directly added to wellsof a microplate that is coated with streptavidin. Subsequently, amixture of biotinylated GLP-1 specific antibody and a horseradishperoxidase (HRP)-conjugated GLP-1 specific antibody is added toeach well. After the first incubation period, a sandwichimmunocomplex of Streptavidin Biotin-Antibody GLP-1(7-36)/(9-36) HRP-conjugated antibody is formed and attached to the wall ofthe plate. The unbound HRP-conjugated antibody is removed in asubsequent washing step. For the detection of this immunocomplex,each well is then incubated with a substrate solution in a timed reactionand then measured in a spectrophotometric microplate reader. Theenzymatic activity of the immunocomplex bound to GLP-1 (7-36)/(9-36)on the wall of the microtiter well is directly proportional to the amount ofTotal GLP-1 in the sample.

Reagent Preparation:

(1) Prior to use allow all reagents to come to room temperature. Reagents from different kit lot numbers should not be combined or interchanged.
(2) ELISA Wash Concentrate must be diluted to working solution prior to use. Please see REAGENTS section for details.
(3) Reconstitute all standards and controls by adding 1.0 mL of deminerialized water to each vial. Allow the standards and controls to sit undisturbed for 10 minutes, and then mix well by gentle vortexing. These reconstituted standards and controls must be stored at - 20 C or below. Do not exceed 3 freeze-thaw cycles.
(4) Test Configuration
(5) Prepare Total GLP-1 Antibody Mixture: mixing Total GLP-1 Tracer Antibody and Total GLP-1 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.

Sample Collection: (1) No special preparation of the individual is necessary prior to specimen collection. However, fasting sample and non-fasting/glucose induced sample may present great significance for Total GLP-1 (7-36)/(9-36) level. (2) EDTA Plasma collection tube such as Vacutainer? BD366643 or similar products must be used for sample collection. (3) Whole blood should be collected into a 10 mL lavender top Vacutainer? EDTA-plasma tube. Process the whole blood to get plasma specimen according to the Vacutainer? manufacturer?s instruction. (4) It is optional to immediately add 100 µL of DPP-4 inhibitor to the collected EDTA whole blood right after the collection (within 30 seconds). Invert tube to mix well and place the tube on ice bath. (5) Plasma samples should be stored at 2 ? 8 C if they will be tested within 3 hours of collection. For longer storage, it is recommended to store the plasma sample at -70 C. Aliquot samples before freezing if necessary. Avoid freeze-thaw until perform this test for sample Total GLP-1 (7-36)/(9-36) measurement.

Assay Procedure:

(1) Place a sufficient number of streptavidin-coated microwell strips/wells in a holder to run Total GLP-1 standards, controls and unknown samples in duplicate.
(2) Add 100 µL of standards, controls and test samples into the designated microwell.
(3) Add 100 µL of Total GLP-1 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 200 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 50 µL of ELISA Stop Solution into each of the wells. Mix gently.
(10) Read the absorbance at wavelength 450nm/620 nm within 10 minutes in a microplate reader.

Calculation of Results:

1. Calculate the average absorbance for each pair of duplicate test results.
   2. Subtract the average absorbance of the STD 1 (0 ng/mL) from the average absorbance of all other readings to obtain the corrected absorbance.
   3. The standard curve is generated by the corrected absorbances of all standard levels on the ordinate against the standard concentration on the abscissa using point-to-point or log-log paper. Appropriate computer-assisted data reduction programs may also be used for the calculation of results. We recommend using Point-to-Point or Quadratic curve fit. The GLP-1 (7-36) concentrations for the controls and test samples are read directly from the standard curve using their respective corrected absorbance

Assay Precision: The intra-assay precision was determined by 8 replicates for 2 control samples in a single assay. The inter-assay precision was determined by 8 individual assays in different dates with 2 control samples.

Restrictions: For Research Use only

Handling

Precaution of Use: The reagents must be used in a professional laboratory environment and are for in vitro diagnostic use only. Source material (e.g. highly purified bovine serum albumin) of bovine serum was derived in the contiguous 48 United States. It was obtained only from healthy donor animals maintained under veterinary supervision and found free of contagious diseases. Wear gloves while performing this assay and handle these reagents as if they are potentially infectious. Avoid contact with reagents containing TMB, hydrogen peroxide, or sulfuric acid. TMB may cause irritation to skin and mucous membranes and cause an allergic skin reaction. TMB is a suspected carcinogen. Sulfuric acid may cause severe irritation on contact with skin. Do not get in eyes, on skin, or on clothing. Do not ingest or inhale fumes. On contact, flush with copious amounts of water for at least 15 minutes. Use Good Laboratory Practices.

Storage: 4 °C

Target Details for GLP-1

Target: GLP-1 (Glucagon-like peptide 1 (GLP-1))

Alternative Name: GLP-1 (GLP-1 Products)

Synonyms: GLP1 ELISA Kit, GLP2 ELISA Kit, GRPP ELISA Kit, GLP-1 ELISA Kit, Glu ELISA Kit, PPG ELISA Kit, glucagon ELISA Kit, GCG ELISA Kit, Gcg ELISA Kit