CLIA 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) 86-93 89
EDTA plasma(n=5) 78-101 83
heparin plasma(n=5) 87-102 91

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% 97-104% 78-104% 81-94%
EDTA plasma(n=5) 95-102% 87-99% 92-99% 90-98%
heparin plasma(n=5) 86-102% 98-105% 92-101% 78-98%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value 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
Nanomaterials An OMV-Based Nanovaccine Confers Safety and Protection against Pathogenic Escherichia coli via Both Humoral and Predominantly Th1 Immune Responses in … Pubmed: 33233490
Diagnostics Systemic Alterations of Immune Response-Related Proteins during Glaucoma Development in the Murine Model DBA/2J Pubmed: 32585848
Scientific Reports The bovine cumulus proteome is influenced by maturation condition and maturational competence of the oocyte Pubmed: 32555221
Int Immunopharmacol Complement component 3 prevents imiquimod-induced psoriatic skin inflammation by inhibiting apoptosis in mice Pubmed: 32535539
Italian Journal of Animal Science Effects of chitooligosaccharide supplementation on laying performance, egg quality, blood biochemistry, antioxidant capacity and immunity of laying hens during the …
Dev Comp Immunol A deep-sea pathogenic Bacillus subtilis isolate employs different strategies to escape the killing of teleost and murine complements 33545212
Animal Effect of alfalfa hay and starter feed supplementation on caecal microbiota and fermentation, growth, and health of yak calves 33610512
British Journal of Pharmacology The suppression of Brd4 inhibits peripheral plasma cell differentiation and exhibits therapeutic potential for systemic lupus erythematosus
Scientific Reports Maternal Serum Proteomic Profiles of Pregnant Women With Type 1 Diabetes 34940989
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