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Multiplex Assay Kit for Obestatin (OB) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-94 84
EDTA plasma(n=5) 92-99 95
heparin plasma(n=5) 85-102 97
sodium citrate plasma(n=5) 84-91 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Obestatin (OB) ,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 Obestatin (OB) ,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 Obestatin (OB) ,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) 91-102% 97-105% 86-99% 78-89%
EDTA plasma(n=5) 96-105% 79-103% 91-105% 85-101%
heparin plasma(n=5) 90-98% 81-96% 80-101% 78-101%
sodium citrate plasma(n=5) 88-101% 88-95% 79-94% 81-105%

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#:OB) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Journal of Obesity and Metabolic Research Evaluation of effects of diet on serum obestatin levels in over-weight patients with polycystic ovary syndrome Org:Source
Turk J Gastroenterol Significance of appetite hormone ghrelin and obestatin levels in the assessment of the severity of acute pancreatitis Pubmed:25141321
Acta Paediatrica Serum complement factor 5a levels are associated with nonalcoholic fatty liver disease in obese children DOI: 10.1111/apa.14106
BMC Endocrine Disorders The role of the adipocytokines vaspin and visfatin in vascular endothelial function and insulin resistance in obese children Pubmed: 31771561
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
CEC039Mu ELISA Kit for Obestatin (OB) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC039Mu Multiplex Assay Kit for Obestatin (OB) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.
KSC039Mu11 ELISA Kit DIY Materials for Obestatin (OB) Main materials for "Do It (ELISA Kit) Yourself".