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親環(huán)素A(CYPA)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

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

  • 親環(huán)素A(CYPA)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 親環(huán)素A(CYPA)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 親環(huán)素A(CYPA)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • SEA979Mu.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過(guò)ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)親環(huán)素A(CYPA),經(jīng)檢測(cè)與其它相似物質(zhì)無(wú)明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來(lái)源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。

回收率

分別于定值血清及血漿樣本中加入一定量的親環(huán)素A(CYPA)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 84-101 95
EDTA plasma(n=5) 88-104 101
heparin plasma(n=5) 97-105 101

精密度

精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
批內(nèi)差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內(nèi)加入適量的親環(huán)素A(CYPA),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中親環(huán)素A(CYPA)含量的測(cè)定值與理論值的比率。

樣本 1:2 1:4 1:8 1:16
serum(n=5) 93-101% 86-101% 94-103% 85-99%
EDTA plasma(n=5) 97-105% 94-102% 97-104% 95-102%
heparin plasma(n=5) 90-104% 91-99% 85-99% 84-98%

穩(wěn)定性

經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來(lái)進(jìn)行操作可減少人為誤差。

實(shí)驗(yàn)流程

1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
2. 加樣(標(biāo)準(zhǔn)品及樣本)100µL,37°C孵育1小時(shí);
3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
4. 洗板3次;
5. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
6. 洗板5次;
7. 加TMB底物90µL,37°C孵育10-20分鐘;
8. 加終止液50µL,立即450nm讀數(shù)。

實(shí)驗(yàn)原理

將親環(huán)素A(CYPA)抗體包被于96孔微孔板中,制成固相載體,向微孔中分別加入標(biāo)準(zhǔn)品或標(biāo)本,其中的親環(huán)素A(CYPA)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的親環(huán)素A(CYPA)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標(biāo)記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過(guò)氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。顏色的深淺和樣品中的親環(huán)素A(CYPA)呈正相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。

相關(guān)產(chǎn)品

編號(hào) 適用物種:Mus musculus (Mouse,小鼠) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
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參考文獻(xiàn)

雜志 參考文獻(xiàn)
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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