Mini Samples ELISA Kit for Complement Component 5a (C5a) Mus musculus (Mouse) Mini ELISA

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  • Mini Samples ELISA Kit for Complement Component 5a (C5a) Packages (Simulation)
  • Mini Samples ELISA Kit for Complement Component 5a (C5a) Packages (Simulation)
  • Mini Samples ELISA Kit for Complement Component 5a (C5a) Results demonstration
  • MEA388Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-98 87
EDTA plasma(n=5) 94-102 98
heparin plasma(n=5) 94-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Complement Component 5a (C5a) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Complement Component 5a (C5a) 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 Mini Samples Complement Component 5a (C5a) 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) 86-101% 79-91% 91-101% 92-101%
EDTA plasma(n=5) 78-95% 88-101% 97-105% 79-97%
heparin plasma(n=5) 98-105% 92-101% 88-95% 85-98%

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 1×60µL Assay Diluent A 1×6mL
Detection Reagent B 1×60µL Assay Diluent B 1×6mL
TMB Substrate 1×4.5mL Stop Solution 1×3mL
Wash Buffer (30 × concentrate) 1×10mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 20µL Stop Solution. Read at 450nm immediately.

Mini Samples ELISA Kit for Complement Component 5a (C5a)

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 Mini Samples Complement Component 5a (C5a). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Complement Component 5a (C5a). 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 Mini Samples Complement Component 5a (C5a), 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 Mini Samples Complement Component 5a (C5a) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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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