Multiplex Assay Kit for Complement Factor D (CFD) ,etc. by FLIA (Flow Luminescence Immunoassay)

CF-D; DF; PFD; ADN; Adipsin; Properdin Factor D; C3 convertase activator

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 94-105 101
EDTA plasma(n=5) 84-103 89
heparin plasma(n=5) 83-97 89
sodium citrate plasma(n=5) 90-98 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Factor D (CFD) ,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 Complement Factor D (CFD) ,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 Complement Factor D (CFD) ,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% 78-105% 91-99% 79-93%
EDTA plasma(n=5) 97-105% 82-92% 95-102% 95-102%
heparin plasma(n=5) 85-93% 85-99% 82-105% 95-105%
sodium citrate plasma(n=5) 81-97% 88-104% 80-94% 93-105%

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#:CFD) 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
Journal of Diabetes Investigation Relationship between serum adipsin and the first phase of glucose‐stimulated insulin secretion in individuals with different glucose tolerance Pubmed:29432659
Endocrine Practice CORRELATION OF INCREASED SERUM ADIPSIN WITH INCREASED CARDIOVASCULAR RISKS IN ADULT PATIENTS WITH GROWTH HORMONE DEFICIENCY Pubmed: 30657359
BMC Endocrine Disorders Association between plasma adipsin level and mild cognitive impairment in Chinese patients with type 2 diabetes: a cross-sectional study Pubmed: 31651303
Scientific Reports HFD-induced hepatic lipid accumulation and inflammation are decreased in Factor D deficient mouse Pubmed: 33067533
Biomedicine & Pharmacotherapy Body fat distribution and circulating adipsin are related to metabolic risks in adult patients with newly diagnosed growth hormone deficiency and improve after … Pubmed: 33254428
Nephrol Dial Transplant Association between thrombotic microangiopathy and activated alternative complement pathway in malignant nephrosclerosis 33367879
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