Multiplex Assay Kit for Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array)
Overview
Properties
- Product No.CMA388Ra
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryInfection immunityImmune moleculeAutoimmunity
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Recovery
Matrices listed below were spiked with certain level of recombinant Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 94-102 | 98 |
| EDTA plasma(n=5) | 79-88 | 85 |
| heparin plasma(n=5) | 95-105 | 98 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) 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 5a (C5a) ,etc. by CBA (Cytometric Bead Array) 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) | 90-98% | 84-98% | 94-102% | 86-99% |
| EDTA plasma(n=5) | 97-105% | 84-101% | 92-99% | 79-103% |
| heparin plasma(n=5) | 84-93% | 80-91% | 98-105% | 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 |
Assay procedure summary
1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
Test principle
Anti-Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads, to make solid phase carrier. Standard or sample were added to the magnetic beads, respectively, in which Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Complement Component 5a (C5a) ,etc. by CBA (Cytometric Bead Array) in the sample and corresponding value of MFI (Median Fluorescence Intensity). Flow Cytometer was used to determine and calculate the sample concentration.
Giveaways
Increment services
Citations
- Cathepsin D is released after severe tissue trauma in vivo and is capable of generating C5a in vitroScienceDirect: S0161589011008297
- Elevated complement factor C5a in maternal and umbilical cord plasma in preeclampsiaPubMed: 23415845
- Complement component 5 contributes to poor disease outcome in humans and mice with pneumococcal meningitisPubMed: PMC3195471
- Efficacy of a synthetic antimicrobial peptidomimetic versus vancomycin in a Staphylococcus epidermidis device-related murine peritonitis modelPubmed: 23645587
- Conversion from external fixator to intramedullary nail causes a second hit and impairs fracture healing in a severe trauma modelPubmed: 23070742
- Therapeutic effects of sesame oil on monosodium urate crystal-induced acute inflammatory response in ratsPubmed: 24353977
- Evaluation of the Markers of Inflammation in the Umbilical Cord Blood of Newborns of Mothers with ThrombophiliaPubmed:25209155
- Complement Split Products C3a/C5a and Receptors: Are They Regulated by Circulating Angiotensin II Type 1 Receptor Autoantibody in Severe Preeclampsia?PubMed: 26485247
- Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitisPubmed:27235358
- Consistency and pathophysiological characterization of a rat polymicrobial sepsis model via the improved cecal ligation and puncture surgeryPubmed:26802602
- Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos10183
- Cadmium-induced immune abnormality is a key pathogenic event in human and rat models of preeclampsiapubmed:27511439
- Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expressionpubmed:29031586
- Serum complement factor 5a levels are associated with nonalcoholic fatty liver disease in obese childrenDOI: 10.1111/apa.14106
- Splenectomy modulates early immuno-inflammatory responses to trauma-hemorrhage and protects mice against secondary sepsisPubmed: 30291296
- The Beneficial Effects of Lactobacillus plantarum PS128 on High-Intensity, Exercise-Induced Oxidative Stress, Inflammation, and Performance in TriathletesPubmed: 30736479
- Intrathecal complement activation by the classical pathway in tick-borne encephalitisPubmed: 30850976
- Sepsis causes right ventricular myocardial inflammation independent of pulmonary hypertension in a porcine sepsis modelPubmed: 31247004
- C5a/C5aR Pathway is Critical for the Pathogenesis of PsoriasisPubmed: 31447855
- Novel evidence that an alternative complement cascade pathway is involved in optimal mobilization of hematopoietic stem/progenitor cells in Nlrp3 …
- C5aR deficiency attenuates the breast cancer development via the p38/p21 axisPubmed: 32669478
- A Novel Evidence That Mannan Binding Lectin (MBL) Pathway of Complement Cascade Activation is Involved in Homing and Engraftment of Hematopoietic Stem …Pubmed: 32406006
- Modulation of C5a-C5aR interactions against murine mammary cancer cell line
- C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney diseasePubmed:35503088
