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

CF-B; BF; BFD; CFAB; GBG; H2-Bf; PBF2; B-Factor; Properdin; Properdin factor B; C3/C5 convertase; Glycine-rich beta glycoprotein

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 94-101 98
EDTA plasma(n=5) 80-101 82
heparin plasma(n=5) 79-99 91
sodium citrate plasma(n=5) 93-102 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Factor B (CFB) ,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 B (CFB) ,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 B (CFB) ,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) 79-96% 95-105% 93-105% 92-101%
EDTA plasma(n=5) 85-101% 85-101% 82-94% 78-93%
heparin plasma(n=5) 81-101% 80-92% 78-89% 85-93%
sodium citrate plasma(n=5) 92-99% 85-99% 79-102% 95-103%

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#:CFB) 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
BMC Nephrology Complement in patients receiving maintenance hemodialysis: functional screening and quantitative analysis BioMed: 14712369
J. Proteome Res. Identification and Confirmation of Differentially Expressed Fucosylated Glycoproteins in the Serum of Ovarian Cancer Patients Using a Lectin Array and LC–MS/MS ACS: pr300330z
Pediatr Int Atypical hemolytic uremic syndrome: Korean pediatric series PubMed: 25443527
Journal of Neuroimmunology Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitis Pubmed:27235358
Biochemical and Biophysical Research Communications Adipose tissue complement factor B promotes adipocyte maturation. pubmed:29137982
Journal of proteome research Serum exosomes from newborn piglets restrict porcine epidemic diarrhea virus infection
journal of human genetics Identification of susceptibility locus shared by IgA nephropathy and inflammatory bowel disease in a Chinese Han population Pubmed: 31857673
Nephrol Dial Transplant Association between thrombotic microangiopathy and activated alternative complement pathway in malignant nephrosclerosis 33367879
JOURNAL OF GENERAL VIROLOGY Changes in complement alternative pathway components, factor B and factor H during dengue virus infection in the AG129 mouse 33410734
J Proteome Res Early Diagnostic Ability of Human Complement Factor B in Pancreatic Cancer Is Partly Linked to Its Potential Tumor-Promoting Role 34766501
J Clin Med A Three-Protein Panel to Support the Diagnosis of Sepsis in Children Pubmed:35329889
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