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Ⅱ型前膠原羧基端原肽(PIICP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Procollagen II C-Terminal Propeptide (PIICP)

P2CP; C-Propeptide Of Type II Procollagen; Procollagen II Carboxy Terminal Propeptide

  • Ⅱ型前膠原羧基端原肽(PIICP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • Ⅱ型前膠原羧基端原肽(PIICP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • Ⅱ型前膠原羧基端原肽(PIICP)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
  • SEA964Hu.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)Ⅱ型前膠原羧基端原肽(PIICP),經(jīng)檢測(cè)與其它相似物質(zhì)無明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來源的限制,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Ⅱ型前膠原羧基端原肽(PIICP)(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) 87-105 98
EDTA plasma(n=5) 89-96 93
heparin plasma(n=5) 94-104 97

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Ⅱ型前膠原羧基端原肽(PIICP),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Ⅱ型前膠原羧基端原肽(PIICP)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) 78-90% 83-102% 80-88% 89-101%
EDTA plasma(n=5) 93-101% 97-104% 78-89% 97-104%
heparin plasma(n=5) 94-101% 99-105% 92-104% 78-96%

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Ⅱ型前膠原羧基端原肽(PIICP)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Ⅱ型前膠原羧基端原肽(PIICP)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的Ⅱ型前膠原羧基端原肽(PIICP)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Ⅱ型前膠原羧基端原肽(PIICP)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。

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參考文獻(xiàn)

雜志 參考文獻(xiàn)
Experimental and Therapeutic Medicine Evaluation of the effect of N-acetyl-glucosamine administration on biomarkers for cartilage metabolism in healthy individuals without symptoms of arthritis: A … [10.3892]
Functional Foods in Health & Disease Evaluation of the effect of administering N-acetyl-glucosamine-containing green tea supplement on biomarkers for cartilage metabolism in healthy individuals without symptoms of arthritis: a randomized double-blind placebo-controlled clinical study [309]
EXPERIMENTAL AND THERAPEUTIC MEDICINE Effect of N-acetylglucosamine administration on cartilage metabolism and safety in healthy subjects without symptoms of arthritis: A case report [pubmed:28413518]
Experimental?and?Therapeutic?Medicine Evaluation of the effect of salmon nasal proteoglycan on biomarkers for cartilage metabolism in individuals with knee joint discomfort: A randomized double?blind placebo?controlled clinical study [etm:14]
Experimental and Therapeutic Medicine Evaluation of the efficacy of Ajuga decumbens extract supplement in individuals with knee discomfort associated with physical activity: A randomized, double?blind, placebo?controlled study [10.3892/etm.2017.5064]
EXPERIMENTAL AND THERAPEUTIC MEDICINE Evaluation of the chondroprotective action of N?acetylglucosamine in a rat experimental osteoarthritis [pubmed:28912864]
Functional Foods In Health And Disease Evaluation of the effect of N-acetyl-glucosamine administration on biomarkers for cartilage metabolism in healthy individuals: a randomized double-blind placebo-controlled clinical study [view/366]
Spine No effects of hyperosmolar culture medium on tissue regeneration by human degenerated nucleus pulposus cells despite upregulation extracellular matrix genes [fulltext:2018/03010]
Advances in?Clinical?and Experimental?Medicine Fibulin-3 and other cartilage metabolism biomarkers in relationship to calprotectin (MRP8/14) and disease activity in rheumatoid arthritis patients treated with?… [68362.pdf]
Functional Food Research サケ鼻軟骨由來プロテオグリカン摂取による関節(jié)保護(hù)効果 []
Spine No Effects of Hyperosmolar Culture Medium on Tissue Regeneration by Human Degenerated [Pubmed: 25856264]
Applied Physiology Nutrition and Metabolism Effectiveness of collagen supplementation on pain scores in healthy individuals with self-reported knee pain; A randomized controlled trial [Pubmed: 31990581]
International Journal of Molecular Sciences Bone Morphogenetic Proteins for Nucleus Pulposus Regeneration [Pubmed: 32295299]
Biomolecules Concerted Actions by PIICP, CTXII, and TNF-|á in Patients with Juvenile Idiopathic Arthritis [33924892]
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