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Human GC(Glucagon) ELISA Kit

Citations(4)Uniprot : P01275

Cat.No.:E-EL-H2237
Reactivity: Human

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96T
48T
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96T*5
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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℃, 12 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), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 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)	96.10	209.10	817.70	86.80	218.30	757.10
Standard deviation	6.30	11.50	33.50	5.20	10.70	39.40
CV (%)	6.56	5.50	4.10	5.99	4.90	5.20

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)	87-102	93
EDTA plasma(n=8)	88-99	94
Cell culture media(n=8)	92-106	99

Linearity

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

Target Information

Database Links	SwissProt: P01275
Synonyms	GCG, 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

DIABETES OBESITY & METABOLISM (2020) IF: 5.9

Effect of fully automated closed-loop insulin delivery using faster aspart versus standard aspart on gluco-regulatory hormones in type 2 diabetes

DOI: 10.1111/dom.14185
PMID: 32902125	Sample: Plasma

Computational and Structural Biotechnology Journal (2020) IF: 6.018

Lipid metabolic signatures deviate in sepsis survivors compared to non-survivors

DOI: 10.1016/j.csbj.2020.11.009
PMID: 33304464	Sample: Blood

Pharmaceutics (2022) IF: 6.525

Endocrine and Metabolic Impact of Oral Ingestion of a Carob-Pod-Derived Natural-Syrup-Containing D-Pinitol: Potential Use as a Novel Sweetener in Diabetes

DOI: 10.3390/pharmaceutics14081594
PMID: 36015220	Sample: Plasma

Nutrients (2022) IF: 6.706

Pharmacokinetics and Endocrine Effects of an Oral Dose of D-Pinitol in Human Fasting Healthy Volunteers

DOI: 10.3390/nu14194094
PMID: 36235746	Sample: Plasma

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