ELISA Kit for Fetuin B (FETUB)

FETU-B; 16G2; Gugu; IRL685; Fetuin-like protein IRL685

Specificity

This assay has high sensitivity and excellent specificity for detection of Fetuin B (FETUB).
No significant cross-reactivity or interference between Fetuin B (FETUB) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Fetuin B (FETUB) and the recovery rates were calculated by comparing the measured value to the expected amount of Fetuin B (FETUB) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-102 92
EDTA plasma(n=5) 88-101 93
heparin plasma(n=5) 84-93 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fetuin B (FETUB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fetuin B (FETUB) 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 Fetuin B (FETUB) 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) 95-105% 85-99% 86-102% 82-89%
EDTA plasma(n=5) 80-88% 87-102% 81-104% 91-103%
heparin plasma(n=5) 94-102% 94-102% 86-103% 78-93%

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
Endocrine Serum fetuin B level increased in subjects of nonalcoholic fatty liver disease: a case-control study pubmed:27628583
Endocrine Increased serum levels of fetuin B in patients with coronary artery disease pubmed:28822077
DIABETES & METABOLISM Plasma fetuin-B concentrations are associated with insulin resistance and first-phase glucose-stimulated insulin secretion in individuals with different degrees of … Pubmed:29514765
Clinical & Investigative Medicine Serum level of fetuin B is associated with osteoporosis: a 4-year prospective study in China Pubmed:29603688
American Journal of Translational Research Fetuin B aggravates liver X receptor-mediated hepatic steatosis through AMPK in HepG2 cells and mice Pubmed: 30972177
Endocrine Connections Increased serum fetuin-B concentration is associated with HOMA-β and indices of liver steatosis in women with polycystic ovary syndrome: a pilot study Pubmed: 31307012
J Transl Med Multi-omic signatures of atherogenic dyslipidaemia: pre-clinical target identification and validation in humans 33407555
J Nephrol Citrate high volume on-line hemodiafiltration modulates serum Interleukin-6 and Klotho levels: the multicenter randomized controlled study ¡°Hephaestus¡± 33559851
Nutrients High Fat, High Sugar Diet and DJOS Bariatric Surgery Influence Plasma Levels of Fetuin-B, Growth Differentiation Factor-15, and Pentraxin 3 in Diet-Induced Obese … 34684637
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