ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9) Mus musculus (Mouse) Sandwich ELISA

MAC; Membrane Attack Complex

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  • ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9) Packages (Simulation)
  • ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9) Packages (Simulation)
  • ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9) Results demonstration
  • SEC350Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 96
EDTA plasma(n=5) 84-101 89
heparin plasma(n=5) 88-95 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Terminal Complement Complex C5b-9 (C5b-9) 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) 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) 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) 80-97% 81-88% 95-102% 87-102%
EDTA plasma(n=5) 88-105% 79-97% 89-96% 95-102%
heparin plasma(n=5) 82-91% 87-101% 99-105% 97-105%

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
TMB Substrate 1×9mL Stop Solution 1×6mL
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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Terminal Complement Complex C5b-9 (C5b-9)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Terminal Complement Complex C5b-9 (C5b-9). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Terminal Complement Complex C5b-9 (C5b-9). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Terminal Complement Complex C5b-9 (C5b-9), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Terminal Complement Complex C5b-9 (C5b-9) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Effect of thiol functionalization on the hemo-compatibility of PLGA nanoparticlesWiley: source
  • Complement component 5 contributes to poor disease outcome in humans and mice with pneumococcal meningitisPubMed: PMC3195471
  • Glucosylated polymeric nanoparticles: A sweetened approach against blood compatibility paradoxScienceDirect: S0927776513001720
  • The effect of normovolemic modified ultrafiltration on inflammatory mediators, endotoxins, terminal complement complexes and clinical outcome in high-risk cardiac surgery patientsPubmed: 23429100
  • The efficacy of recombinant human soluble thrombomodulin for the treatment of shiga toxin associated hemolytic uremic syndrome model micePubmed:25694534
  • Adjuvant treatment with dexamethasone plus anti-C5 antibodies improves outcome of experimental pneumococcal meningitis: a randomized controlled trialPubMed: 26272468
  • Thrombin‐activatable fibrinolysis inhibitor influences disease severity in humans and mice with pneumococcal meningitisPubMed: 26340319
  • Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos10183
  • Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitisPMC5234106
  • Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expressionpubmed:29031586
  • Effects of immunoadsorption combined with membrane filtration on complement markers–Results of a randomized, controlled, crossover study
  • Complement factor H contributes to mortality in humans and mice with bacterial meningitisPubmed: 31883521
  • C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney diseasePubmed:35503088

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