CLIA Kit for Complement Component 4d (C4d) Homo sapiens (Human) Sandwich CLIA

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  • CLIA Kit for Complement Component 4d (C4d) Packages (Simulation)
  • CLIA Kit for Complement Component 4d (C4d) Packages (Simulation)
  • CLIA Kit for Complement Component 4d (C4d) Results demonstration
  • SCS103Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 4d (C4d) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 4d (C4d) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-104 99
EDTA plasma(n=5) 89-101 96
heparin plasma(n=5) 78-95 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 4d (C4d) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 4d (C4d) 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 Component 4d (C4d) 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) 83-90% 88-102% 84-102% 80-91%
EDTA plasma(n=5) 94-104% 95-104% 78-91% 85-95%
heparin plasma(n=5) 93-101% 89-96% 88-102% 80-93%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

CLIA Kit for Complement Component 4d (C4d)

Test principle

The microplate provided in this kit has been pre-coated with an antibody specific to Complement Component 4d (C4d). Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to Complement Component 4d (C4d). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Complement Component 4d (C4d) level in the sample or standard.;

Citations

  • Measurement of Serum Levels of Complement Component 4d as a Quantitative Test of Antibody-Mediated RejectionContent: 144

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