ELISA Kit for Complement Factor B (CFB) Mus musculus (Mouse) Sandwich ELISA

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

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  • ELISA Kit for Complement Factor B (CFB) Packages (Simulation)
  • ELISA Kit for Complement Factor B (CFB) Packages (Simulation)
  • ELISA Kit for Complement Factor B (CFB) Results demonstration
  • SEC011Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-99 89
EDTA plasma(n=5) 81-101 92
heparin plasma(n=5) 85-93 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 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 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 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) 96-104% 91-101% 80-105% 84-103%
EDTA plasma(n=5) 90-99% 79-93% 91-103% 89-99%
heparin plasma(n=5) 80-105% 96-104% 95-102% 78-101%

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
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.

ELISA Kit for Complement Factor B (CFB)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Complement Factor B (CFB). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Complement Factor B (CFB). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Complement Factor B (CFB), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Complement Factor B (CFB) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Complement in patients receiving maintenance hemodialysis: functional screening and quantitative analysisBioMed: 14712369
  • Identification and Confirmation of Differentially Expressed Fucosylated Glycoproteins in the Serum of Ovarian Cancer Patients Using a Lectin Array and LC–MS/MSACS: pr300330z
  • Atypical hemolytic uremic syndrome: Korean pediatric seriesPubMed: 25443527
  • Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitisPubmed:27235358
  • Adipose tissue complement factor B promotes adipocyte maturation.pubmed:29137982
  • Serum exosomes from newborn piglets restrict porcine epidemic diarrhea virus infection
  • Identification of susceptibility locus shared by IgA nephropathy and inflammatory bowel disease in a Chinese Han populationPubmed: 31857673
  • Association between thrombotic microangiopathy and activated alternative complement pathway in malignant nephrosclerosis33367879
  • Changes in complement alternative pathway components, factor B and factor H during dengue virus infection in the AG129 mouse33410734
  • Early Diagnostic Ability of Human Complement Factor B in Pancreatic Cancer Is Partly Linked to Its Potential Tumor-Promoting Role34766501
  • A Three-Protein Panel to Support the Diagnosis of Sepsis in ChildrenPubmed:35329889

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