ELISA Kit for Complement Component 3a (C3a)

Specificity

This assay has high sensitivity and excellent specificity for detection of Complement Component 3a (C3a).
No significant cross-reactivity or interference between Complement Component 3a (C3a) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 3a (C3a) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 3a (C3a) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-95 89
EDTA plasma(n=5) 82-104 92
heparin plasma(n=5) 84-102 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 3a (C3a) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 3a (C3a) 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 3a (C3a) 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) 81-93% 95-105% 78-96% 95-105%
EDTA plasma(n=5) 80-92% 92-102% 86-94% 86-103%
heparin plasma(n=5) 97-104% 89-103% 79-101% 96-104%

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
22 Immobilization of soluble complement receptor 1 on islets. ScienceDirect: S0142961211002596
Journal of Reproductive Immunology Elevated complement factor C5a in maternal and umbilical cord plasma in preeclampsia PubMed: 23415845
Journal of Antimicrobial Chemotherapy Efficacy of a synthetic antimicrobial peptidomimetic versus vancomycin in a Staphylococcus epidermidis device-related murine peritonitis model Pubmed: 23645587
SpringerPlus Therapeutic effects of sesame oil on monosodium urate crystal-induced acute inflammatory response in rats Pubmed: 24353977
Gynecol Obstet Invest Complement Split Products C3a/C5a and Receptors: Are They Regulated by Circulating Angiotensin II Type 1 Receptor Autoantibody in Severe Preeclampsia? PubMed: 26485247
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
Journal of Neuroimmunology Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitis Pubmed:27235358
Plos One Alternative Pathway Inhibition by Exogenous Factor H Fails to Attenuate Inflammation and Vascular Leakage in Experimental Pneumococcal Sepsis in Mice Pubmed:26872035
Digital Repository Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos 10183
Colloids Surf B Biointerfaces. Interaction of gold and silver nanoparticles with human plasma: Analysis of protein corona reveals specific binding patterns. pubmed:28131092
Pediatric Pulmonology Accuracy of complement activation product levels to detect infected pleural effusion in rats pubmed:28474458
Journal of Immunology The Complement C3a–C3aR Axis Promotes Development of Thoracic Aortic Dissection via Regulation of MMP2 Expression Pubmed:29367209
Artificial Cells Nanomedicine and Biotechnology Journal Long term safety and immunological effects of a nanobiotherapeutic, bovine poly-[hemoglobin-catalase-superoxide dismutase-carbonic anhydrase], after four weekly … Pubmed:29873524
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