Multiplex Assay Kit for Growth Differentiation Factor 5 (GDF5) ,etc. by CBA (Cytometric Bead Array)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-93 84
EDTA plasma(n=5) 90-104 93
heparin plasma(n=5) 91-101 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Growth Differentiation Factor 5 (GDF5) ,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 Growth Differentiation Factor 5 (GDF5) ,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 Growth Differentiation Factor 5 (GDF5) ,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) 96-104% 80-95% 98-105% 83-92%
EDTA plasma(n=5) 89-101% 82-101% 93-101% 89-96%
heparin plasma(n=5) 79-94% 78-90% 79-94% 84-101%

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

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 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 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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