Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

CPAMD4; C5a anaphylatoxin; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 4

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

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  • Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMB969Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-105 99
EDTA plasma(n=5) 87-105 92
heparin plasma(n=5) 80-91 80
sodium citrate plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 5 (C5) ,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 Component 5 (C5) ,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 Component 5 (C5) ,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) 94-101% 86-103% 82-93% 96-105%
EDTA plasma(n=5) 91-101% 91-105% 89-103% 93-101%
heparin plasma(n=5) 92-102% 89-101% 94-102% 79-92%
sodium citrate plasma(n=5) 93-101% 95-102% 94-102% 84-95%

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#:C5) 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.

Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease.PubMed: 22941511
  • Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitisPubmed:27235358
  • Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expressionpubmed:29031586
  • iTRAQ‐Based Proteomic Analysis Reveals Protein Profile in Plasma from Children with AutismPubmed:29274201
  • Impaired insulin signaling in the B10.D2-Hc H2 H2-T18/oSnJ mouse model of complement factor 5 (C5) deficiencyPubmed: 31084499
  • Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancyPubmed: 31162828
  • C5aR deficiency attenuates the breast cancer development via the p38/p21 axisPubmed: 32669478

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