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Mouse GC(Glucagon) ELISA Kit

Citations(6)Uniprot : P55095

Cat.No.:E-EL-M0555
Reactivity: Mouse

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96T
48T
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96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	31.25-2000 pg/mL
Sensitivity	18.75 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes GC in samples. No significant cross-reactivity or interference between GC and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of GC concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with GC. During the reaction, GC in the sample or standard competes with a fixed amount of GC on the solid phase supporter for sites on the Biotinylated Detection Ab specific to GC. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of GC in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level GC were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level GC were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	108.40	253.50	739.40	98.90	254.20	733.70
Standard deviation	5.60	12.70	31.80	6.60	14.20	32.30
CV (%)	5.17	5.01	4.30	6.67	5.59	4.40

Recovery

The recovery of GC spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	93-110	100
EDTA plasma(n=8)	92-102	97
Cell culture media(n=8)	87-100	92

Linearity

Samples were spiked with high concentrations of Mouse GC and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P55095
Synonyms	GCG, GLP1, GLP2, GRPP
Research Area	Cancer, Metabolism, Developmental biology, Signal transduction, Stem cells

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

CELL (2023) IF: 66.849
Light modulates glucose metabolism by a retina-hypothalamus-brown adipose tissue axis

DOI: 10.1016/j.cell.2022.12.024

PMID: 36669474
Sample: Serum
DIABETES (2016) IF: 8.7840
Insulin-like growth factor binding protein (IGFBP)-1 could through its RGD domain improve glucose regulation and insulin sensitivity

DOI: 10.2337/db16-0997

Sample: Plasma
Frontiers in Nutrition (2022) IF: 6.576
Slowly Digestible Carbohydrate Diet Ameliorates Hyperglycemia and Hyperlipidemia in High-Fat Diet/Streptozocin-Induced Diabetic Mice

DOI: 10.3389/fnut.2022.854725

PMID: 35495933
Sample: liver tissue
ENDOCRINOLOGY (2023) IF: 5.051
Aspirin suppresses hepatic glucagon signaling through decreasing production of thromboxane A2

DOI: 10.1210/endocr/bqac217

Sample: plasma,liver
Journal of Functional Foods (2022) IF: 5.223
Arjunolic acid from Cyclocarya paliurus selectively inhibits glucagon secretion from α cells and ameliorates diabetes via ephrin-A1 and EphA4 interaction

DOI: 10.1016/j.jff.2022.105323

Sample: serum
NEUROPEPTIDES (2020) IF: 3.286
GLP-1 preserves β cell function via improvement on islet insulin signaling in high fat diet feeding mice

DOI: 10.1016/j.npep.2020.102110

PMID: 33307381
Sample: serum

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