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Multiplex Assay Kit for Obestatin (OB) ,etc. by CBA (Cytometric Bead Array)

Specificity

This assay has high sensitivity and excellent specificity for detection of Obestatin (OB) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Obestatin (OB) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Obestatin (OB) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Obestatin (OB) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-103 99
EDTA plasma(n=5) 83-94 88
heparin plasma(n=5) 89-98 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Obestatin (OB) ,etc. by CBA (Cytometric Bead Array) 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 CBA (Cytometric Bead Array) 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 CBA (Cytometric Bead Array) 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) 80-91% 78-101% 95-103% 96-105%
EDTA plasma(n=5) 80-95% 92-102% 99-105% 92-102%
heparin plasma(n=5) 78-103% 92-99% 83-94% 84-97%

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. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 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.
CMC039Mu Multiplex Assay Kit for Obestatin (OB) ,etc. by CBA (Cytometric Bead Array) FLIA Kit 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".