ELISA Kit for Complement Component 3 (C3)

ASP; CPAMD1; Complement Protein C3; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1; C3a anaphylatoxin; Acylation stimulating protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Complement Component 3 (C3).
No significant cross-reactivity or interference between Complement Component 3 (C3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 3 (C3) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 3 (C3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-95 91
EDTA plasma(n=5) 95-104 98
heparin plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 3 (C3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 3 (C3) 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 3 (C3) 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) 98-105% 79-104% 90-98% 88-101%
EDTA plasma(n=5) 80-101% 95-105% 94-101% 90-104%
heparin plasma(n=5) 83-101% 78-101% 97-105% 98-105%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Artificial Organs Experimental Research With Synthetic Copolymer-Coated Cardiopulmonary Bypass Circuits: Inflammatory and Thrombogenicity Analysis Wiley: source
Journal of Clinical Immunology Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease. PubMed: 22941511
Lipids in Health and Disease Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteome PubMed: PMC3480889
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
Veterinary Immunology and Immunopathology Oral administration of heat-inactivated Mycobacterium bovis reduces the response of farmed red deer to avian and bovine tuberculin Pubmed:27032499
International Journal of Molecular Sciences Identification of Serum Peptidome Signatures of Non-Small Cell Lung Cancer Pubmed:27043541
Food Chem. Comparative proteomics of milk fat globule membrane in goat colostrum and mature milk Pubmed:27173528
Scientific Reports Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting. pubmed:28694465
Veterinary Microbiology The response of red deer to oral administration of heat-inactivated Mycobacterium bovis and challenge with a field strain. pubmed:28888638
Proteomics Clinical Applications iTRAQ‐Based Proteomic Analysis Reveals Protein Profile in Plasma from Children with Autism Pubmed:29274201
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