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

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Complement Component 3a (C3a) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

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) 90-103 96
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 93-101 96

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) 82-95% 93-101% 91-98% 80-99%
EDTA plasma(n=5) 96-105% 89-96% 93-101% 80-101%
heparin plasma(n=5) 80-90% 82-105% 84-91% 83-94%

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.

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Magazine Citations
22 Immobilization of soluble complement receptor 1 on islets. ScienceDirect: S0142961211002596
Journal of Reproductive Immunology Elevated complement factor C5a in maternal and umbilical cord plasma in preeclampsia PubMed: 23415845
Journal of Antimicrobial Chemotherapy Efficacy of a synthetic antimicrobial peptidomimetic versus vancomycin in a Staphylococcus epidermidis device-related murine peritonitis model Pubmed: 23645587
SpringerPlus Therapeutic effects of sesame oil on monosodium urate crystal-induced acute inflammatory response in rats Pubmed: 24353977
Gynecol Obstet Invest Complement Split Products C3a/C5a and Receptors: Are They Regulated by Circulating Angiotensin II Type 1 Receptor Autoantibody in Severe Preeclampsia? PubMed: 26485247
Am J Reprod Immunol Human IgG Antinuclear Antibodies Induce Pregnancy Loss in Mice by Increasing Immune Complex Deposition in Placental Tissue: In Vivo Study PubMed: 26388133
Journal of Neuroimmunology Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitis Pubmed:27235358
Plos One Alternative Pathway Inhibition by Exogenous Factor H Fails to Attenuate Inflammation and Vascular Leakage in Experimental Pneumococcal Sepsis in Mice Pubmed:26872035
Digital Repository Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos 10183
Colloids Surf B Biointerfaces. Interaction of gold and silver nanoparticles with human plasma: Analysis of protein corona reveals specific binding patterns. pubmed:28131092
Pediatric Pulmonology Accuracy of complement activation product levels to detect infected pleural effusion in rats pubmed:28474458
Journal of Immunology The Complement C3a–C3aR Axis Promotes Development of Thoracic Aortic Dissection via Regulation of MMP2 Expression Pubmed:29367209
Artificial Cells Nanomedicine and Biotechnology Journal Long term safety and immunological effects of a nanobiotherapeutic, bovine poly-[hemoglobin-catalase-superoxide dismutase-carbonic anhydrase], after four weekly … Pubmed:29873524
Cytotherapy Mesenchymal stromal cell secretory factors induce sustained improvements in islet function pre-and post-transplantation Doi: 10.1016/j.jcyt.2018.07.007
Journal of NeuroVirology Intrathecal complement activation by the classical pathway in tick-borne encephalitis Pubmed: 30850976
Clinical Immunology Pathogenic roles of anti-C1q antibodies in recurrent pregnancy loss
Materials Science and Engineering: C Hemocompatibility of biodegradable Zn-0.8 wt%(Cu, Mn, Li) alloys
Nanomedicine Complement-mediated hypersensitivity reactions to an amphotericin B-containing lipid complex (Abelcet) in pediatric patients and anesthetized rats: Benefits of slow infusion 33549818
EXPERIMENTAL EYE RESEARCH Increased ratios of complement factors C3a to C3 in aqueous humor and serum mark glaucoma progression 33493474
FASEB J C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease Pubmed:35503088
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