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M2-型丙酮酸激酶(PKM2)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for Pyruvate kinase isozymes M2 (PKM2)

M2PK; PKM; M2-PK; PKM1; CTHBP; OIP3; PK3; PKM; TCB; THBP1; Pyruvate kinase muscle isozyme; Thyroid hormone-binding protein 1; Cytosolic thyroid hormone-binding protein; Pyruvate Kinase, Muscle

  • M2-型丙酮酸激酶(PKM2)檢測試劑盒(酶聯(lián)免疫吸附試驗法)產(chǎn)品包裝(模擬)
  • M2-型丙酮酸激酶(PKM2)檢測試劑盒(酶聯(lián)免疫吸附試驗法)產(chǎn)品包裝(模擬)
  • M2-型丙酮酸激酶(PKM2)檢測試劑盒(酶聯(lián)免疫吸附試驗法)實驗結(jié)果圖
  • SEA588Hu.jpg標準曲線圖
  • Certificate通過ISO 9001、ISO 13485質(zhì)量體系認證

特異性

本試劑盒用于檢測M2-型丙酮酸激酶(PKM2),經(jīng)檢測與其它相似物質(zhì)無明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來源的限制,不可能完成對所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測,因此本試劑盒有可能與未經(jīng)檢測的其它物質(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M2-型丙酮酸激酶(PKM2)(加標樣品),重復(fù)測定并計算其均值,回收率為測定值與理論值的比率。

樣本回收率范圍(%)平均回收率(%)
serum(n=5)80-8985
EDTA plasma(n=5)81-9690
heparin plasma(n=5)91-10595

精密度

精密度用樣品測定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對低、中、高值定值樣本進行定量檢測,每份樣本連續(xù)測定20 次,分別計算不同濃度樣本的平均值及SD值。
批間差:選取3個不同批次的試劑盒分別對低、中、高值定值樣本進行定量測定,每個樣本使用同一試劑盒重復(fù)測定8次,分別計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M2-型丙酮酸激酶(PKM2),并倍比稀釋成1:2,1:4,1:8,1:16的待測樣本,線性范圍即為稀釋后樣本中M2-型丙酮酸激酶(PKM2)含量的測定值與理論值的比率。

樣本1:21:41:81:16
serum(n=5)88-96%78-105%79-97%87-101%
EDTA plasma(n=5)95-105%99-105%90-103%92-101%
heparin plasma(n=5)88-97%81-99%94-103%93-102%

穩(wěn)定性

經(jīng)測定,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%。
為減小外部因素對試劑盒破壞前后檢測值的影響,實驗室的環(huán)境條件需盡量保持一致,尤其是實驗室內(nèi)溫度、濕度及溫育條件。其次由同一實驗員來進行操作可減少人為誤差。

實驗流程

1. 實驗前標準品、試劑及樣本的準備;
2. 加樣(標準品及樣本)100µL,37°C孵育1小時;
3. 吸棄,加檢測溶液A100µL,37°C孵育1小時;
4. 洗板3次;
5. 加檢測溶液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)。

實驗原理

將M2-型丙酮酸激酶(PKM2)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準品或標本,其中的M2-型丙酮酸激酶(PKM2)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的M2-型丙酮酸激酶(PKM2)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標記的親和素,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M2-型丙酮酸激酶(PKM2)呈正相關(guān)。用酶標儀在450nm波長下測定吸光度(O.D.值),計算樣品濃度。

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編號適用物種:Homo sapiens (Human,人)應(yīng)用(僅供研究使用,不用于臨床診斷!)
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參考文獻

雜志參考文獻
Cancer ResearchAntigen Presentation by Dendritic Cells in Tumors Is Disrupted by Altered Metabolism that Involves Pyruvate Kinase M2 and Its Interaction with SOCS3[AACR: 70189]
Molecular BioSystemsAbnormal levels of heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1) in tumour tissue and blood samples from patients diagnosed with lung cancer[Pubmed:25483567]
Tumour BiolM2 isoform of pyruvate kinase (PKM2) is upregulated in Kazakh’s ESCC and promotes proliferation and migration of ESCC cells[PubMed: 26404132]
OncotargetIdentification of a serum biomarker panel for the differential diagnosis of cholangiocarcinoma and primary sclerosing cholangitis[Pubmed:29707118]
Journal of?NeurotraumaActeoside improves muscle atrophy and motor function by inducing new myokine secretion in chronic spinal cord injury[Pubmed: 30318996]
Cell ReportsSecreted Pyruvate Kinase M2 Promotes Lung Cancer Metastasis through Activating the Integrin Beta1/FAK Signaling Pathway[Pubmed: 32049010]
Chrysin serves as a novel inhibitor of DGKα/FAK interaction to suppress the malignancy of esophageal squamous cell carcinoma (ESCC)[]
Cancer Cell InternationalBufalin Induced Mitochondrial Dysfunction Promotes Apoptosis of Glioma Cells by Regulating Annexin A2 and DRP1 Proteins[]
Cancer Cell IntBufalin induces mitochondrial dysfunction and promotes apoptosis of glioma cells by regulating Annexin A2 and DRP1 protein expression[34376212]
Life (Basel)Putative Association between Low Baseline Gene Expression in the Peripheral Blood and Clinical Remission in Rheumatoid Arthritis Patients Treated with Tofacitinib[34947916]