Multiplex Assay Kit for Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) Mus musculus (Mouse) Multiplex ELISA

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA388Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-101 90
EDTA plasma(n=5) 96-105 102
heparin plasma(n=5) 79-101 89
sodium citrate plasma(n=5) 80-89 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 FLIA (Flow Luminescence Immunoassay) 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 FLIA (Flow Luminescence Immunoassay) 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% 81-102% 79-94% 89-97%
EDTA plasma(n=5) 80-90% 82-89% 81-96% 81-98%
heparin plasma(n=5) 90-102% 79-99% 79-97% 81-93%
sodium citrate plasma(n=5) 89-96% 91-99% 89-103% 81-94%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:C5a) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Complement Component 5a (C5a) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

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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

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