Mini Samples ELISA Kit for Complement Component 5 (C5) Mus musculus (Mouse) Mini ELISA

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

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  • Mini Samples ELISA Kit for Complement Component 5 (C5) Packages (Simulation)
  • Mini Samples ELISA Kit for Complement Component 5 (C5) Packages (Simulation)
  • Mini Samples ELISA Kit for Complement Component 5 (C5) Results demonstration
  • MEB969Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Mini Samples Complement Component 5 (C5) and the recovery rates were calculated by comparing the measured value to the expected amount of Mini Samples Complement Component 5 (C5) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
EDTA plasma(n=5) 83-90 86
heparin plasma(n=5) 89-102 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Complement Component 5 (C5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Complement Component 5 (C5) 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 Mini Samples Complement Component 5 (C5) 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% 83-91% 80-94% 93-105%
EDTA plasma(n=5) 94-101% 81-101% 83-104% 81-104%
heparin plasma(n=5) 94-101% 94-101% 79-94% 81-90%

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 1×60µL Assay Diluent A 1×6mL
Detection Reagent B 1×60µL Assay Diluent B 1×6mL
TMB Substrate 1×4.5mL Stop Solution 1×3mL
Wash Buffer (30 × concentrate) 1×10mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 20µL Stop Solution. Read at 450nm immediately.

Mini Samples ELISA Kit for Complement Component 5 (C5)

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 Mini Samples Complement Component 5 (C5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Complement Component 5 (C5). 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 Mini Samples Complement Component 5 (C5), 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 Mini Samples Complement Component 5 (C5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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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