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Rat GLUT4(Glucose Transporter 4) ELISA Kit

Cat.No.:E-EL-R0430
Reactivity: Rat

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.63-40 ng/mL
Sensitivity	0.38 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat GLUT4 in samples. No significant cross-reactivity or interference between Rat GLUT4 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 Rat GLUT4 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat GLUT4. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat GLUT4 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat GLUT4, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat GLUT4. You can calculate the concentration of Rat GLUT4 in the samples 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°C
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 Rat GLUT4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat GLUT4 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 (ng/mL)	2.03	4.50	19.63	2.21	4.26	19.21
Standard deviation	0.11	0.21	0.98	0.13	0.18	0.75
CV (%)	5.42	4.67	4.99	5.88	4.23	3.90

Recovery

The recovery of Rat GLUT4 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)	91-101	96
EDTA plasma (n=8)	88-101	94
Cell culture media (n=8)	83-98	90

Linearity

Samples were spiked with high concentrations of Rat GLUT4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	SLC2A4, Solute Carrier Family 2 Member 4(Facilitated Glucose Transporter)
Research Area	Cancer, Cardiovascular, Metabolism, Signal transduction, Stem cells

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Frontiers in Endocrinology (2022) IF: 5.555
Programming With Varying Dietary Fat Content Alters Cardiac Insulin Receptor, Glut4 and FoxO1 Immunoreactivity in Neonatal Rats, Whereas High Fat Programming Alters Cebpa Gene Expression in Neonatal Female Rats

DOI: 10.3389/fendo.2021.772095

PMID: 35069436
Sample: hearts
ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY (2020) IF: 3.292
Chronic exposure of bisphenol A impairs carbohydrate and lipid metabolism by altering corresponding enzymatic and metabolic pathways

DOI: 10.1016/j.etap.2020.103387

PMID: 32339907
Sample: Serum
RSC Advances (2021) IF: 3.361
Metabolomic profiling to reveal the therapeutic potency of Posidonia oceanica nanoparticles in diabetic rats

DOI: 10.1039/D0RA09606G

Sample: Serum
BIOMEDICINE & PHARMACOTHERAPY (2018) IF: 3.457
Raspberry ketone and Garcinia Cambogia rebalanced disrupted insulin resistance and leptin signaling in rats fed high fat fructose diet

DOI: 10.1016/j.biopha.2018.11.079

PMID: 30530230
Sample: Tissue homogenate
BIOMEDICINE & PHARMACOTHERAPY (2018) IF: 3.457
Balanites aegyptiaca ameliorates insulin secretion and decreases pancreatic apoptosis in diabetic rats: Role of SAPK/JNK pathway

DOI: 10.1016/j.biopha.2018.03.167

Sample: Plasma,Tissue homogenate
JOURNAL OF ETHNOPHARMACOLOGY (2020) IF: 3.69
Upregulation of MC4R and PPAR-α expression mediates the anti-obesity activity of Moringa oleifera Lam. in high-fat diet-induced obesity in rats

DOI: 10.1016/j.jep.2020.112541

PMID: 31911179
Sample: Tissue homogenate
Frontiers in Pharmacology (2018) IF: 3.831
Novel Trajectories of Bromocriptine Antidiabetic Action: Leptin-IL-6/ JAK2/p-STAT3/SOCS3, p-IR/p-AKT/GLUT4, PPAR-γ/Adiponectin, Nrf2/PARP-1, and GLP-1

DOI: 10.3389/fphar.2018.00771

Sample: Serum
Scientific African (2021)
Adverse effects of perinatal protein restriction on glucose homeostasis in offspring of Sprague-Dawley rats

DOI: 10.1016/j.sciaf.2021.e01036

Sample: gastrocnemius muscle

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