Wide-range ELISA Kit for Complement Factor B (CFB)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Wide-range Complement Factor B (CFB).
No significant cross-reactivity or interference between Wide-range Complement Factor B (CFB) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Wide-range Complement Factor B (CFB) and the recovery rates were calculated by comparing the measured value to the expected amount of Wide-range Complement Factor B (CFB) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-104 101
EDTA plasma(n=5) 79-102 83
heparin plasma(n=5) 86-102 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wide-range Complement Factor B (CFB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Wide-range Complement Factor B (CFB) 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 Wide-range Complement Factor B (CFB) 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) 90-102% 92-105% 82-90% 82-90%
EDTA plasma(n=5) 88-104% 84-101% 92-101% 78-98%
heparin plasma(n=5) 78-89% 80-102% 97-105% 98-105%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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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