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ELISA Kit for Retinol Binding Protein 4, Plasma (RBP4)

PRBP; RBP; Plasma retinol-binding protein

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 99
EDTA plasma(n=5) 84-99 90
heparin plasma(n=5) 92-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Retinol Binding Protein 4, Plasma (RBP4) 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, Plasma (RBP4) 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, Plasma (RBP4) 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) 92-101% 83-104% 96-104% 96-105%
EDTA plasma(n=5) 80-102% 90-98% 81-97% 87-94%
heparin plasma(n=5) 84-92% 95-105% 78-91% 92-99%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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
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