Multiplex Assay Kit for Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) Oryctolagus cuniculus (Rabbit) Multiplex ELISA

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

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  • Multiplex Assay Kit for Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA387Rb.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-101 91
EDTA plasma(n=5) 79-97 89
heparin plasma(n=5) 78-95 82
sodium citrate plasma(n=5) 95-105 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 3a (C3a) ,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 3a (C3a) ,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 3a (C3a) ,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) 97-104% 81-94% 94-102% 78-93%
EDTA plasma(n=5) 86-103% 81-91% 78-93% 79-103%
heparin plasma(n=5) 93-101% 93-101% 88-99% 88-96%
sodium citrate plasma(n=5) 82-104% 85-103% 82-96% 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#:C3a) 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 3a (C3a) ,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.

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Citations

  • Immobilization of soluble complement receptor 1 on islets.ScienceDirect: S0142961211002596
  • Elevated complement factor C5a in maternal and umbilical cord plasma in preeclampsiaPubMed: 23415845
  • Efficacy of a synthetic antimicrobial peptidomimetic versus vancomycin in a Staphylococcus epidermidis device-related murine peritonitis modelPubmed: 23645587
  • Therapeutic effects of sesame oil on monosodium urate crystal-induced acute inflammatory response in ratsPubmed: 24353977
  • Complement Split Products C3a/C5a and Receptors: Are They Regulated by Circulating Angiotensin II Type 1 Receptor Autoantibody in Severe Preeclampsia?PubMed: 26485247
  • Human IgG Antinuclear Antibodies Induce Pregnancy Loss in Mice by Increasing Immune Complex Deposition in Placental Tissue: In Vivo StudyPubMed: 26388133
  • Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitisPubmed:27235358
  • Alternative Pathway Inhibition by Exogenous Factor H Fails to Attenuate Inflammation and Vascular Leakage in Experimental Pneumococcal Sepsis in MicePubmed:26872035
  • Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos10183
  • Interaction of gold and silver nanoparticles with human plasma: Analysis of protein corona reveals specific binding patterns.pubmed:28131092
  • Accuracy of complement activation product levels to detect infected pleural effusion in ratspubmed:28474458
  • The Complement C3a–C3aR Axis Promotes Development of Thoracic Aortic Dissection via Regulation of MMP2 ExpressionPubmed:29367209
  • Long term safety and immunological effects of a nanobiotherapeutic, bovine poly-[hemoglobin-catalase-superoxide dismutase-carbonic anhydrase], after four weekly …Pubmed:29873524
  • Mesenchymal stromal cell secretory factors induce sustained improvements in islet function pre-and post-transplantationDoi: 10.1016/j.jcyt.2018.07.007
  • Intrathecal complement activation by the classical pathway in tick-borne encephalitisPubmed: 30850976
  • Pathogenic roles of anti-C1q antibodies in recurrent pregnancy loss
  • Hemocompatibility of biodegradable Zn-0.8 wt%(Cu, Mn, Li) alloys
  • Complement-mediated hypersensitivity reactions to an amphotericin B-containing lipid complex (Abelcet) in pediatric patients and anesthetized rats: Benefits of slow infusion33549818
  • Increased ratios of complement factors C3a to C3 in aqueous humor and serum mark glaucoma progression33493474
  • C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney diseasePubmed:35503088

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