ELISA Kit for Cyclophilin A (CYPA)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Cyclophilin A (CYPA).
No significant cross-reactivity or interference between Cyclophilin A (CYPA) and analogues was observed.

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) 81-92 88
EDTA plasma(n=5) 92-101 98
heparin plasma(n=5) 92-99 96

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) 88-103% 85-99% 88-104% 98-105%
EDTA plasma(n=5) 80-101% 91-105% 92-101% 96-105%
heparin plasma(n=5) 89-104% 89-103% 97-105% 78-91%

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Atherosclerosis EMMPRIN and its ligand cyclophilin A regulate MT1-MMP, MMP-9 and M-CSF during foam cell formation PubMed: 19758589
Biomarkers Cyclophilin-A: a novel biomarker for untreated male essential hypertension Pubmed: 24164594
Cardiovascular Diabetology Plasma level of cyclophilin A is increased in patients with type 2 diabetes mellitus and suggests presence of vascular disease Pubmed:24502618
International Journal of Biological Sciences 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 Infarction Pubmed:25552928
Medicine Cyclophilin A in Ruptured Intracranial Aneurysm PubMed: 26426668
Mol Cell Biochem PPIA rs6850: A> G single-nucleotide polymorphism is associated with raised plasma cyclophilin A levels in patients with coronary artery disease PubMed: 26702934
Medicine (Baltimore) Urinary Cyclophilin A as a New Marker for Diabetic Nephropathy: A Cross-Sectional Analysis of Diabetes Mellitus PubMed: 26496315
International Journal of Cardiology Cyclophilin A modulates bone marrow-derived CD117(+) cells and enhances ischemia-induced angiogenesisvia the SDF-1/CXCR4 axis. Pubmed:27057951
Clinica Chimica Acta Novel findings of secreted cyclophilin A in diabetic nephropathy and its association with renal protection of dipeptidyl peptidase 4 inhibitor pubmed:27823952
Cardiovascular Diabetology 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
Clinical Science Metformin attenuates effects of cyclophilin A on macrophages, reduces lipid uptake and secretion of cytokines by repressing decreased AMPK activity Pubmed:29382697
Journal of Molecular Histology Cyclophilin a increases CD68 cell infiltration in rat experimental periodontitis Pubmed:29318411
Free Radical Research Cyclophilin A and CD147 associate with progression of diabetic nephropathy Pubmed: 30572748
The Kaohsiung Journal of Medical Sciences Association of cyclophilin A level and pulse pressure in predicting recurrence of cerebral infarction Pubmed: 31670477
Cells. Cyclophilin A/EMMPRIN Axis Is Involved in Pro-Fibrotic Processes Associated with Thoracic Aortic Aneurysm of Marfan Syndrome Patients Pubmed: 31936351
JAMA Neurology APPENDIX A: Relationship between cyclophilin A levels and matrix metalloproteinase-9 activity in cerebrospinal fluid of cognitively normal apolipoprotein E4 carriers … Pubmed: 24030206
Menoufia Medical Journal Urinary podocalyxin and cyclophilin A: markers for early detection of type 2 diabetic nephropathy
Toxins Evaluation of the Therapeutic Effects of Protocatechuic Aldehyde in Diabetic Nephropathy 34437430
Cell Death & Disease Proteomic analysis reveals USP7 as a novel regulator of palmitic acid-induced hepatocellular carcinoma cell death Pubmed:35732625
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