Multiplex Assay Kit for Retinol Binding Protein 4 (RBP4) ,etc. by FLIA (Flow Luminescence Immunoassay)

PRBP; RBP; Plasma retinol-binding protein; Retinol Binding Protein 4, Plasma

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Retinol Binding Protein 4 (RBP4) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Retinol Binding Protein 4 (RBP4) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Retinol Binding Protein 4 (RBP4) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Retinol Binding Protein 4 (RBP4) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 80-91 88
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Retinol Binding Protein 4 (RBP4) ,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 Retinol Binding Protein 4 (RBP4) ,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 Retinol Binding Protein 4 (RBP4) ,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) 91-99% 80-96% 93-101% 78-101%
EDTA plasma(n=5) 90-99% 84-101% 86-101% 93-103%
heparin plasma(n=5) 79-99% 81-101% 86-104% 80-99%

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#:RBP4) 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
Diabetes Technology & Therapeutics Proteomic Identification of Human Urinary Biomarkers in Diabetes Mellitus Type 2 Liebert: 20100078
Journal of Pharmaceutical and Biomedical Analysis Effect of high dose thiamine on the levels of urinary protein biomarkers in diabetes mellitus type 2 PubMed: 21130593
Evid Based Complement Alternat Med Quercetin Protects against Cadmium-Induced Renal Uric Acid Transport System Alteration and Lipid Metabolism Disorder in Rats PubMed: PMC3368504
Journal of Endocrinology Combined galanin with insulin improves insulin sensitivity of diabetic rat muscles Pubmed:24501381
J Endocrinol Central alarin ameliorated insulin resistance of adipocytes in type 2 diabetic rats Pubmed:25240061
Peptides Effect of endogenous galanin on glucose transporter 4 expression in cardiac muscle of type 2 diabetic rats Pubmed:25445608
Obesity Surgery Long-term Effect of Ileal Transposition on Adipokine Serum Level in Zucker (Orl)-Lepr fa Fatty Rats Pubmed:25697126
Clin Exp Nephrol Differentially expressed urinary biomarkers in children with idiopathic nephrotic syndrome PubMed: 26351173
Experimental and Therapeutic Medicine Expression of retinol binding protein 4 and nuclear factor-ÎşB in diabetic rats with atherosclerosis and the intervention effect of pioglitazone Pubmed:27446311
Experimental and Therapeutic Medicine Expression of retinol binding protein 4 and nuclear factor-κB in diabetic rats with atherosclerosisand the intervention effect of pioglitazone. pubmed:27446311
Journal of Chromatography A Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosis S0021967317311858
BMC Cancer Screening for immune-potentiating antigens from hepatocellular carcinoma patients after radiofrequency ablation by serum proteomic analysis Pubmed:29386009
Nutrition & Diabetes Lack of pronounced changes in the expression of fatty acid handling proteins in adipose tissue and plasma of morbidly obese humans Pubmed:29335416
Molecular Medicine Reports RBP4 regulates trophoblastic cell proliferation and invasion via the PI3K/AKT signaling pathway Pubmed:30015949
Relationship between retinol and risk of diabetic retinopathy: a case-control study
Scientific Reports Retinol binding protein 4 abundance in plasma and tissues is related to body fat deposition in cattle Pubmed: 31147589
Potassium supplementation blunts the effects of high salt intake on serum retinol‐binding protein 4 levels in healthy individuals
Docosahexaenoic acid (DHA), a nutritional supplement, modulates steroid insensitivity in asthma
Eur J Nutr The dietary inflammatory index is associated with anti-and pro-inflammatory adipokines in Brazilian schoolchildren 33575861
Eur J Pediatr Pro-and anti-inflammatory adipokines are associated with cardiometabolic risk markers in Brazilian schoolchildren 33834274
Sci Rep Ileal transposition helps to regulate plasma hepatokine levels in obese Zucker (Crl: ZUC (ORL)-Lepr fa) rats 33833309
Int Arch Occup Environ Health Biomarkers of cadmium exposure and renal function in estuarine adult villagers 34773507
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