Multiplex Assay Kit for Cyclophilin A (CYPA) ,etc. by FLIA (Flow Luminescence Immunoassay) Mus musculus (Mouse) Multiplex ELISA

PPIA; CyP-A; CYPH; Peptidylprolyl Isomerase A; Peptidyl-Prolyl Isomerase A; Peptidyl-prolyl cis-trans isomerase A; Cyclosporin A-binding protein

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

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  • Multiplex Assay Kit for Cyclophilin A (CYPA) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Cyclophilin A (CYPA) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Cyclophilin A (CYPA) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA979Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Cyclophilin A (CYPA) and the recovery rates were calculated by comparing the measured value to the expected amount of Cyclophilin A (CYPA) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-105 91
EDTA plasma(n=5) 85-96 89
heparin plasma(n=5) 82-90 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cyclophilin A (CYPA) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cyclophilin A (CYPA) 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 Cyclophilin A (CYPA) 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) 92-99% 91-101% 90-102% 87-95%
EDTA plasma(n=5) 92-101% 80-99% 86-103% 93-105%
heparin plasma(n=5) 92-99% 98-105% 83-95% 88-99%

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#:CYPA) 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. 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.

Multiplex Assay Kit for Cyclophilin A (CYPA) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Anti-Cyclophilin A (CYPA) 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 Cyclophilin A (CYPA) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Cyclophilin A (CYPA) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Cyclophilin A (CYPA) in the sample and corresponding value of MFI (Median Fluorescence Intensity).

Giveaways

Citations

  • EMMPRIN and its ligand cyclophilin A regulate MT1-MMP, MMP-9 and M-CSF during foam cell formationPubMed: 19758589
  • Cyclophilin-A: a novel biomarker for untreated male essential hypertensionPubmed: 24164594
  • Plasma level of cyclophilin A is increased in patients with type 2 diabetes mellitus and suggests presence of vascular diseasePubmed:24502618
  • Decrease in Plasma Cyclophilin A Concentration at 1 Month after Myocardial Infarction Predicts Better Left Ventricular Performance and Synchronicity at 6 Months: A Pilot Study in Patients with ST Elevation Myocardial InfarctionPubmed:25552928
  • Cyclophilin A in Ruptured Intracranial AneurysmPubMed: 26426668
  • PPIA rs6850: A> G single-nucleotide polymorphism is associated with raised plasma cyclophilin A levels in patients with coronary artery diseasePubMed: 26702934
  • Urinary Cyclophilin A as a New Marker for Diabetic Nephropathy: A Cross-Sectional Analysis of Diabetes MellitusPubMed: 26496315
  • Cyclophilin A modulates bone marrow-derived CD117(+) cells and enhances ischemia-induced angiogenesisvia the SDF-1/CXCR4 axis.Pubmed:27057951
  • Novel findings of secreted cyclophilin A in diabetic nephropathy and its association with renal protection of dipeptidyl peptidase 4 inhibitorpubmed:27823952
  • Cyclophilin A enhances macrophage differentiation and lipid uptake in high glucose conditions: a cellular mechanism for accelerated macro vascular disease in diabetes mellitus.pubmed:27809851
  • Metformin attenuates effects of cyclophilin A on macrophages, reduces lipid uptake and secretion of cytokines by repressing decreased AMPK activityPubmed:29382697
  • Cyclophilin a increases CD68 cell infiltration in rat experimental periodontitisPubmed:29318411
  • Cyclophilin A and CD147 associate with progression of diabetic nephropathyPubmed: 30572748
  • Association of cyclophilin A level and pulse pressure in predicting recurrence of cerebral infarctionPubmed: 31670477
  • Cyclophilin A/EMMPRIN Axis Is Involved in Pro-Fibrotic Processes Associated with Thoracic Aortic Aneurysm of Marfan Syndrome PatientsPubmed: 31936351
  • APPENDIX A: Relationship between cyclophilin A levels and matrix metalloproteinase-9 activity in cerebrospinal fluid of cognitively normal apolipoprotein E4 carriers …Pubmed: 24030206
  • Urinary podocalyxin and cyclophilin A: markers for early detection of type 2 diabetic nephropathy
  • Evaluation of the Therapeutic Effects of Protocatechuic Aldehyde in Diabetic Nephropathy34437430
  • Proteomic analysis reveals USP7 as a novel regulator of palmitic acid-induced hepatocellular carcinoma cell deathPubmed:35732625

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