Multiplex Assay Kit for Growth Differentiation Factor 5 (GDF5) ,etc. by FLIA (Flow Luminescence Immunoassay)

CDMP1; LAP4; SYNS2; BMP14; Radotermin; Cartilage-Derived Morphogenetic Protein-1; Bone morphogenetic protein 14; Lipopolysaccharide-associated protein 4

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Growth Differentiation Factor 5 (GDF5) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Growth Differentiation Factor 5 (GDF5) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Growth Differentiation Factor 5 (GDF5) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Growth Differentiation Factor 5 (GDF5) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-105 101
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 96-104 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Growth Differentiation Factor 5 (GDF5) ,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 Growth Differentiation Factor 5 (GDF5) ,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 Growth Differentiation Factor 5 (GDF5) ,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) 93-105% 98-105% 87-97% 91-99%
EDTA plasma(n=5) 89-103% 92-104% 98-105% 85-98%
heparin plasma(n=5) 97-105% 82-103% 90-105% 91-99%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:GDF5) 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 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Carbohydrate Polymers ZrO2 surface chemically coated with hyaluronic acid hydrogel loading GDF-5 for osteogenesis in dentistry PubMed: 23218279
Bone Photo-cured hyaluronic acid-based hydrogels containing growth and differentiation factor 5 (GDF-5) for bone tissue regeneration ScienceDirect: S875632821300481X
J Nanosci Nanotechnol In Vitro Osteogenic Differentiation Enhanced by Zirconia Coated with Nano-Layered Growth and Differentiation Factor-5 Pubmed:27398455
Macromolecular Research Osteoblastic Differentiation of Functionalized Biphasic Hydroxyapatite and b-Tricalcium Phosphate with Recombinant Human Growth and Differentiation (rhGDF-5)
Pharmaceutics 3D Biomimetic Scaffold for Growth Factor Controlled Delivery: An In-Vitro Study of Tenogenic Events on Wharton's Jelly Mesenchymal Stem Cells 34575523
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