ELISA Kit for Complement Component 5 (C5)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Complement Component 5 (C5).
No significant cross-reactivity or interference between Complement Component 5 (C5) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 84-97 90
heparin plasma(n=5) 90-103 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 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 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 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) 88-103% 94-101% 89-96% 84-96%
EDTA plasma(n=5) 90-98% 82-92% 91-98% 83-101%
heparin plasma(n=5) 92-105% 92-105% 84-92% 88-102%

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.

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Magazine Citations
Journal of Clinical Immunology Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease. PubMed: 22941511
Journal of Neuroimmunology Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitis Pubmed:27235358
Cancer Letters Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expression pubmed:29031586
Proteomics Clinical Applications iTRAQ‐Based Proteomic Analysis Reveals Protein Profile in Plasma from Children with Autism Pubmed:29274201
American journal of physiology. Endocrinology and metabolism Impaired insulin signaling in the B10.D2-Hc H2 H2-T18/oSnJ mouse model of complement factor 5 (C5) deficiency Pubmed: 31084499
Proteomics Clinical Applications Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancy Pubmed: 31162828
Aging C5aR deficiency attenuates the breast cancer development via the p38/p21 axis Pubmed: 32669478
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
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