Multiplex Assay Kit for Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay)

SLC2A4; Solute carrier family 2, facilitated glucose transporter member 4; Glucose transporter type 4, insulin-responsive

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-97 92
EDTA plasma(n=5) 94-101 97
heparin plasma(n=5) 89-98 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Glucose Transporter 4 (GLUT4) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 82-101% 80-90% 80-97% 86-101%
EDTA plasma(n=5) 81-104% 80-96% 98-105% 96-103%
heparin plasma(n=5) 91-101% 80-95% 91-98% 82-102%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:GLUT4) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Polar Record Glucose transporter-4 in white blood cells of young and old sled dogs: a model for human biomarker development Cambridge: Source
The International Journal of Biochemistry &amp; Cell Biology Conditioning causes an increase in glucose transporter-4 levels in mononuclear cells in sled dogs Pubmed:25236492
Psychoneuroendocrinology Maternal stress predicts altered biogenesis and the profile of mitochondrial proteins in the frontal cortex and hippocampus of adult offspring rats PubMed: 26143539
PLOS ONE Antidiabetic Effect of Galantamine: Novel Effect for a Known Centrally Acting Drug PubMed: 26262991
American Journal of Animal and Veterinary Sciences Metabolic Features of Heart Failure with Different Etiology ofsp11075
Journal of Pharmaceutical Negative Results Hibiscus sabdariffa linn fruits methanolic extract and fractions mediated glucose uptake stimulation and glucose transporter 4 regulation Doi: 10.4103/jpnr.JPNR_14_18
eureka herba indonesia Effect of Noni Fruit Extract (Morinda citrifolia) on Glucose Intake to Diabetes Mellitus White Rat Muscle Tissue
eureka herba indonesia The Effects of Sambiloto Leaf Extract (Andrographis peniculata) on Blood Sugar Regulation: an In Vivo Study
eureka herba indonesia Effect of Bay Leaf Extract (Syzygium polyanthum) on Blood Sugar Regulation via GLUT4 Protein Regulation in White Rat Muscle Tissue Induced Aloxan
Eureka Herba Indonesia Potential Combination of Tinospora crispa, Andrographis paniculata, Cinamomum burmanii, Syzygium polyanthum and Momordica charantia Extracts Against?¡­
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