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

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

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  • Multiplex Assay Kit for Obestatin (OB) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Obestatin (OB) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Obestatin (OB) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMC039Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-101 86
EDTA plasma(n=5) 78-96 87
heparin plasma(n=5) 95-102 99
sodium citrate plasma(n=5) 82-92 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Obestatin (OB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Obestatin (OB) 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) 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) 81-101% 80-92% 95-105% 98-105%
EDTA plasma(n=5) 79-89% 85-101% 78-89% 78-95%
heparin plasma(n=5) 80-99% 78-90% 84-98% 79-101%
sodium citrate plasma(n=5) 80-90% 91-98% 88-102% 89-96%

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.

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

Test principle

Anti-Obestatin (OB) antibody was coated on the surface of magnetic beads. A competitive inhibition reaction is launched between biotin labeled Obestatin (OB) and unlabeled Obestatin (OB) (Standards or samples) with the pre-coated antibody specific to Obestatin (OB). PE-labeled avidin was added. The amount of bound PE conjugate is reverse proportional to the concentration of Obestatin (OB) in the sample, the intensity of MFI developed is reverse proportional to the concentration of Obestatin (OB) in the sample.

Giveaways

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Citations

  • Evaluation of effects of diet on serum obestatin levels in over-weight patients with polycystic ovary syndromeOrg:Source
  • Significance of appetite hormone ghrelin and obestatin levels in the assessment of the severity of acute pancreatitisPubmed:25141321
  • Serum complement factor 5a levels are associated with nonalcoholic fatty liver disease in obese childrenDOI: 10.1111/apa.14106
  • The role of the adipocytokines vaspin and visfatin in vascular endothelial function and insulin resistance in obese childrenPubmed: 31771561

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