ELISA Kit for Complement Component 3 (C3) Sus scrofa; Porcine (Pig) Sandwich ELISA

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

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  • ELISA Kit for Complement Component 3 (C3) Packages (Simulation)
  • ELISA Kit for Complement Component 3 (C3) Packages (Simulation)
  • ELISA Kit for Complement Component 3 (C3) Results demonstration
  • SEA861Po.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 92-101 95
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 78-94 88

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) 99-105% 98-105% 82-90% 91-98%
EDTA plasma(n=5) 89-102% 80-94% 87-101% 78-90%
heparin plasma(n=5) 89-99% 89-103% 91-98% 82-103%

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 Complement Component 3 (C3)

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 Complement Component 3 (C3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Complement Component 3 (C3). 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 Complement Component 3 (C3), 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 Complement Component 3 (C3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Experimental Research With Synthetic Copolymer-Coated Cardiopulmonary Bypass Circuits: Inflammatory and Thrombogenicity AnalysisWiley: source
  • Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease.PubMed: 22941511
  • Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteomePubMed: PMC3480889
  • Human IgG Antinuclear Antibodies Induce Pregnancy Loss in Mice by Increasing Immune Complex Deposition in Placental Tissue: In Vivo StudyPubMed: 26388133
  • Oral administration of heat-inactivated Mycobacterium bovis reduces the response of farmed red deer to avian and bovine tuberculinPubmed:27032499
  • Identification of Serum Peptidome Signatures of Non-Small Cell Lung CancerPubmed:27043541
  • Comparative proteomics of milk fat globule membrane in goat colostrum and mature milkPubmed:27173528
  • Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting. pubmed:28694465
  • The response of red deer to oral administration of heat-inactivated Mycobacterium bovis and challenge with a field strain.pubmed:28888638
  • iTRAQ‐Based Proteomic Analysis Reveals Protein Profile in Plasma from Children with AutismPubmed:29274201
  • An OMV-Based Nanovaccine Confers Safety and Protection against Pathogenic Escherichia coli via Both Humoral and Predominantly Th1 Immune Responses in …Pubmed: 33233490
  • Systemic Alterations of Immune Response-Related Proteins during Glaucoma Development in the Murine Model DBA/2JPubmed: 32585848
  • The bovine cumulus proteome is influenced by maturation condition and maturational competence of the oocytePubmed: 32555221
  • Complement component 3 prevents imiquimod-induced psoriatic skin inflammation by inhibiting apoptosis in micePubmed: 32535539
  • Effects of chitooligosaccharide supplementation on laying performance, egg quality, blood biochemistry, antioxidant capacity and immunity of laying hens during the …
  • A deep-sea pathogenic Bacillus subtilis isolate employs different strategies to escape the killing of teleost and murine complements33545212
  • Effect of alfalfa hay and starter feed supplementation on caecal microbiota and fermentation, growth, and health of yak calves33610512
  • The suppression of Brd4 inhibits peripheral plasma cell differentiation and exhibits therapeutic potential for systemic lupus erythematosus
  • Maternal Serum Proteomic Profiles of Pregnant Women With Type 1 Diabetes34940989

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