Multiplex Assay Kit for Terminal Complement Complex C5b-9 (C5b-9) ,etc. by FLIA (Flow Luminescence Immunoassay)

MAC; Membrane Attack Complex

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Terminal Complement Complex C5b-9 (C5b-9) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Terminal Complement Complex C5b-9 (C5b-9) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Terminal Complement Complex C5b-9 (C5b-9) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Terminal Complement Complex C5b-9 (C5b-9) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-105 99
EDTA plasma(n=5) 83-101 95
heparin plasma(n=5) 78-102 92
sodium citrate plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Terminal Complement Complex C5b-9 (C5b-9) ,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 Terminal Complement Complex C5b-9 (C5b-9) ,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 Terminal Complement Complex C5b-9 (C5b-9) ,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) 79-90% 90-102% 82-102% 99-105%
EDTA plasma(n=5) 84-91% 79-93% 98-105% 87-102%
heparin plasma(n=5) 79-88% 93-101% 84-92% 87-94%
sodium citrate plasma(n=5) 83-102% 96-105% 86-94% 87-99%

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#:C5b-9) 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
Journal of Biomedical Materials Research Part A Effect of thiol functionalization on the hemo-compatibility of PLGA nanoparticles Wiley: source
The Journal of Clinical Investigation Complement component 5 contributes to poor disease outcome in humans and mice with pneumococcal meningitis PubMed: PMC3195471
Colloids and Surfaces B: Biointerfaces Glucosylated polymeric nanoparticles: A sweetened approach against blood compatibility paradox ScienceDirect: S0927776513001720
Perfusion The effect of normovolemic modified ultrafiltration on inflammatory mediators, endotoxins, terminal complement complexes and clinical outcome in high-risk cardiac surgery patients Pubmed: 23429100
Nephrology Dialysis Transplantation The efficacy of recombinant human soluble thrombomodulin for the treatment of shiga toxin associated hemolytic uremic syndrome model mice Pubmed:25694534
journal of neuroinflammation Adjuvant treatment with dexamethasone plus anti-C5 antibodies improves outcome of experimental pneumococcal meningitis: a randomized controlled trial PubMed: 26272468
J Thromb Haemost Thrombin‐activatable fibrinolysis inhibitor influences disease severity in humans and mice with pneumococcal meningitis PubMed: 26340319
Digital Repository Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos 10183
J Neuroinflammation.  Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitis PMC5234106
Cancer Letters Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expression pubmed:29031586
Effects of immunoadsorption combined with membrane filtration on complement markers–Results of a randomized, controlled, crossover study
Journal of Neuroinflammation Complement factor H contributes to mortality in humans and mice with bacterial meningitis Pubmed: 31883521
FASEB J C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease Pubmed:35503088
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
SEC350Hu ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC350Hu Multiplex Assay Kit for Terminal Complement Complex C5b-9 (C5b-9) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.