Multiplex Assay Kit for Platelet Derived Growth Factor C (PDGFC) ,etc. by FLIA (Flow Luminescence Immunoassay)

PDGF-C; SCDGF; VEGF-E; Fallotein; Spinal cord-derived growth factor; Platelet-derived growth factor C, latent form; Platelet-derived growth factor C, receptor-binding form

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-104 87
EDTA plasma(n=5) 91-99 94
heparin plasma(n=5) 99-105 102
sodium citrate plasma(n=5) 87-95 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Platelet Derived Growth Factor C (PDGFC) ,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 Platelet Derived Growth Factor C (PDGFC) ,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 Platelet Derived Growth Factor C (PDGFC) ,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) 89-103% 88-96% 86-96% 89-98%
EDTA plasma(n=5) 85-99% 78-93% 85-96% 90-98%
heparin plasma(n=5) 95-103% 90-98% 96-103% 79-97%
sodium citrate plasma(n=5) 96-104% 92-102% 99-105% 81-90%

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#:PDGFC) 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
Immunobiology Platelet derived growth factor-CC secreted by M2 macrophages induces alpha-smooth muscle actin expression by dermal and gingival fibroblasts PubMed: 23182716
Physiological Reports High glucose induces platelet‐derived growth factor‐C via carbohydrate response element‐binding protein in glomerular mesangial cells Pubmed:27033449
Respiratory Research  Changes in pulmonary endothelial cell properties during bleomycin-induced pulmonary fibrosis Pubmed:29940932
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