Description |
The Mouse glucagon-like peptide-1 (GLP-1) ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of mouse GLP-1in many biological fluids, including serum, plasma, or tissue homogenates. GLP-1 is a gut-derived peptide secreted from intestinal L-cells following a meal. The action of GLP-1 is through the binding of a specific GLP-1 receptor, which is expressed in various tissues, including pancreatic beta cells, pancreatic ducts, and gastric mucosa. GLP-1 exerts its main effect by enhancing glucose-dependent insulin production. GLP-1 has also been demonstrated to delay gastric emptying, suppress inappropriate post-meal glucagon release, and increase satiety.
This assay employs the Sandwich-ELISA technique, in which GLP-1 in the samples or standards is sandwiched between pre-coated an anti-mouse GLP-1 and Biotin-labeled GLP-1 antibody. The Ag-Ab-Ag immune complex is labeled with HRP-avidin and then develops a color reaction after the addition of the TMB substrate. The intensity of the color is positively correlated to the amount of GLP-1 in the sample. The kit has a detection range of 62.5 ng/mL-4000 ng/mL, excellent sensitivity, high specificity, good linearity, precision low than 10%, high recovery, good lot-to-lot consistency. Get access to more details from the product instructions.
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Target Name |
glucagon-like peptide-1,GLP-1 |
Alternative Names |
N/A |
Abbreviation |
GLP-1 |
Species |
Mus musculus (Mouse) |
Sample Types |
serum, plasma, tissue homogenates |
Detection Range |
0.312 pg/mL-20 pg/mL |
Sensitivity |
0.078 pg/mL |
Assay Time |
1-5h |
Sample Volume |
50-100ul |
Detection Wavelength |
450 nm |
Research Area |
Metabolism
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Assay Principle |
quantitative |
Measurement |
Sandwich |
Precision |
Intra-assay Precision (Precision within an assay): CV%<8% | | | | Three samples of known concentration were tested twenty times on one plate to assess. | | Inter-assay Precision (Precision between assays): CV%<10% | | | | Three samples of known concentration were tested in twenty assays to assess. | | | | | | | | | |
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Linearity |
To assess the linearity of the assay, samples were spiked with high concentrations of mouse GLP-1 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. | | Sample | Serum(n=4) | | 1:100 | Average % | 97 | | Range % | 92-103 | | 1:200 | Average % | 92 | | Range % | 85-99 | | 1:400 | Average % | 90 | | Range % | 88-94 | | 1:800 | Average % | 101 | | Range % | 96-105 | |
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Recovery |
The recovery of mouse GLP-1 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. | |
Sample Type | Average % Recovery | Range | | Serum (n=5) | 93 | 87-95 | | EDTA plasma (n=4) | 106 | 103-111 | | | | | | | | | | | | | | | |
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Typical Data |
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. | |
pg/ml | OD1 | OD2 | Average | Corrected | | 20 | 2.401 | 2.552 | 2.477 | 2.344 | | 10 | 2.033 | 2.076 | 2.055 | 1.922 | | 5 | 1.524 | 1.604 | 1.564 | 1.431 | | 2.5 | 1.011 | 1.009 | 1.010 | 0.877 | | 1.25 | 0.524 | 0.527 | 0.526 | 0.393 | | 0.625 | 0.319 | 0.345 | 0.332 | 0.199 | | 0.312 | 0.235 | 0.229 | 0.232 | 0.099 | | 0 | 0.130 | 0.135 | 0.133 | | |
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Materials provided |
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-mouse GLP-1 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled GLP-1 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
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Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution
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Troubleshooting
and FAQs |
ELISA kit FAQs |
Storage |
Store at 2-8°C. Please refer to protocol. |
Lead Time |
3-5 working days |