Multiplex Assay Kit for P-Selectin (SELP) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

CD62P; Selectin, Platelet; PSEL; GMP140; LECAM3; GRMP; GMRP; PADGEM; Leukocyte-endothelial cell adhesion molecule 3; Platelet activation dependent granule-external membrane

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

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  • Multiplex Assay Kit for P-Selectin (SELP) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for P-Selectin (SELP) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for P-Selectin (SELP) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA569Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant P-Selectin (SELP) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of P-Selectin (SELP) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-105 101
EDTA plasma(n=5) 84-101 89
heparin plasma(n=5) 93-104 101
sodium citrate plasma(n=5) 78-101 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level P-Selectin (SELP) ,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 P-Selectin (SELP) ,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 P-Selectin (SELP) ,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) 96-104% 84-92% 91-105% 95-105%
EDTA plasma(n=5) 84-99% 80-90% 80-90% 80-103%
heparin plasma(n=5) 89-102% 85-94% 85-101% 85-99%
sodium citrate plasma(n=5) 80-97% 79-97% 91-99% 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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:SELP) 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 P-Selectin (SELP) ,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

Citations

  • Effect of Hirulog-Like Peptide on Balloon Catheter Injury-Induced Neointimal Formation in Femoral Arteries of Minipigs and Relationship with Inflammatory MediatorsKarger: 000257340
  • Secretome of apoptotic peripheral blood cells (APOSEC) attenuates microvascular obstruction in a porcine closed chest reperfused acute myocardial infarction model: role of platelet aggregation and vasodilationPubMed: 22899170
  • Endothelial gene expression and molecular changes in response to radiosurgery in in vitro and in vivo models of cerebral arteriovenous malformationsPubmed: 24199192
  • Involvement of nitric oxide with activation of Toll-like receptor 4 signaling in mice with dextran sodium sulfate-induced colitisPubmed:24992835
  • MRI-compatible Nb–60Ta–2Zr alloy used for vascular stents: Haemocompatibility and its correlation with protein adsorptionPubmed:25063132
  • Chemical sympathectomy attenuates inflammation, glycocalyx shedding and coagulation disorders in rats with acute traumatic coagulopathy.Pubmed:25325345
  • Systemic and Flap Inflammatory Response Associates with Thrombosis in Flap Venous CrisisPubmed:25448261
  • Effects of glucagon-like peptide-1 on advanced glycation endproduct-induced aortic endothelial dysfunction in streptozotocin-induced diabetic rats: possible roles of Rho kinase- and AMP kinase-mediated nuclear factor κB signaling pathwaysPubmed:26758998
  • Protective effects of sinomenine against LPS-induced inflammation in piglets.pubmed:28757275
  • L-Carnitine Supplementation Increases Trimethylamine-N-Oxide but not Markers of Atherosclerosis in Healthy Aged WomenPubmed: 30485835
  • Evaluating the Platelet Activation Related to the Degradation of Biomaterials by Scheme of Molecular Markers
  • Molecular mechanisms of lead-induced changes of selenium status in mice livers through interacting with selenoprotein P
  • Feasibility study of use of rabbit blood to evaluate platelet activation by medical devicesPubmed: 31838449
  • Polycaprolactone vascular graft with epigallocatechin gallate embedded sandwiched layer-by-layer functionalization for enhanced antithrombogenicity and anti …Pubmed: 31982435
  • The Inhibition of P-Selectin Reduced Severe Acute Lung Injury in Immunocompromised MicePubmed: 32377309
  • The binding of autotaxin to integrins mediates hyperhomocysteinemia-potentiated platelet activation and thrombosis34559203
  • The effect of ChAdOx1 nCov-19 vaccine on arterial thrombosis development and platelet aggregation in female ratsPubmed:35183388
  • The inhibition of TRIM35-mediated TIGAR ubiquitination enhances mitochondrial fusion and alleviates renal ischemia-reperfusion injuryPubmed:35421414
  • Construction of a rabbit model with vinorelbine administration via peripherally inserted central catheter and dynamic monitoring of changes in phlebitis and thrombosisPubmed:35126715

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