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可羅索(KL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Klotho (KL)

KLa; Klotho Alpha

  • 可羅索(KL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 可羅索(KL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 可羅索(KL)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
  • SEH757Mu.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)可羅索(KL),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可羅索(KL)(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) 80-97 85
EDTA plasma(n=5) 96-104 101
heparin plasma(n=5) 80-103 90

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)加入適量的可羅索(KL),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可羅索(KL)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) 92-101% 88-97% 79-101% 96-103%
EDTA plasma(n=5) 88-102% 92-101% 78-105% 89-97%
heparin plasma(n=5) 89-96% 89-96% 78-98% 78-95%

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

將可羅索(KL)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,其中的可羅索(KL)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的可羅索(KL)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可羅索(KL)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!)
RPH757Mu03 可羅索(KL)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
RPH757Mu02 可羅索(KL)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
RPH757Mu01 可羅索(KL)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAH757Mu02 可羅索(KL)多克隆抗體 WB; IHC; ICC; IP.
PAH757Mu01 可羅索(KL)多克隆抗體 WB; IHC; ICC; IP.
LAH757Mu71 可羅索(KL)多克隆抗體(生物素標(biāo)記) WB; IHC; ICC.
LAH757Mu81 可羅索(KL)多克隆抗體(異硫氰酸熒光素標(biāo)記) WB; IHC; ICC; IF.
SEH757Mu 可羅索(KL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
WEH757Mu 可羅索(KL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,寬范圍) Enzyme-linked immunosorbent assay for Antigen Detection.
SCH757Mu 可羅索(KL)檢測(cè)試劑盒(化學(xué)發(fā)光免疫分析法) Chemiluminescent immunoassay for Antigen Detection.
LMH757Mu 可羅索(KL)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.
KSH757Mu01 可羅索(KL)檢測(cè)試劑盒DIY材料(酶聯(lián)免疫吸附試驗(yàn)法) 自己動(dòng)手制作ELISA試劑盒的主要材料

參考文獻(xiàn)

雜志 參考文獻(xiàn)
Life Sciences High phosphate feeding induced arterial medial calcification in uremic rats: Roles of Lanthanum carbonate on protecting vasculature [Pubmed: 24012609]
EXPERIMENTAL &amp; CLINICAL CARDIOLOGY Sick Sinus Syndrome May Attribute to the Decrease of ?±-klotho Protein in Mice [Com:Source]
Int J Clin Exp Pathol.? α-Klotho is an acute phase protein and altered by restraint stress in mice [Pubmed:Pmc4203207]
Annales de Biologie Clinique Les biomarqueurs des calcifications vasculaires [PubMed: 26069069]
Mol Med Rep Effect of secondary hyperparathyroidism serum on endothelial cells and intervention with Klotho [PubMed: 25873020]
Calcif Tissue Int Human Preosteoblastic Cell Culture from a Patient with Severe Tumoral Calcinosis-Hyperphosphatemia Due to a NewGALNT3 [PubMed: 25899975]
中華內(nèi)分泌代謝雜志 抗衰老蛋白Klotho 阻斷高糖培養(yǎng)的大鼠腎小球系膜細(xì)胞TLR4/NF-κB p65/NGAL 通路的激活及其作用機(jī)制 [Cn: Article]
Mol Cell Endocrinol The function of miR-199a-5p/Klotho regulating TLR4/NF-κB p65/NGAL pathways in rat mesangial cells cultured with high glucose and the mechanism [PubMed: 26419931]
Arch Med Sci Soluble Klotho protein as a novel serum biomarker in patients with acromegaly [pmc:PMC4754356]
Am J Nephrol Klotho?Deficiency?Aggravates?Tacrolimus-Induced?Renal?Injury?via?the?Phosphatidylinositol?3-Kinase-Akt-Forkhead?Box?Protein?O?Pathway [Pubmed:27174564]
Sci Rep. Klotho preservation via histone deacetylase inhibition attenuates chronic kidney disease-associated bone injury in mice. [pubmed:28387374]
Nephrology Is there a role for newer biomarkers in chronic kidney disease-mineral and bone disorder management? [pubmed:28429560]
Journal of Integrative Nephrology & Andrology Associations between serum fibroblast growth factor 23 level and intradialytic hypotension in hemodialysis patients [issn:2394-2916]
International Journal of Clinical and Experimental Pathology Protective effect of restructuring klotho protein combined with trimetazidine on myocardial injury in mice [ISSN:1936-2625]
Journal of veterinary medical science Changes of Klotho protein and Klotho mRNA expression in a hydroxy-L-proline induced hyperoxaluric rat model [pubmed:28943597]
Cellular Physiology and Biochemistry Klotho Reduces Necroptosis by Targeting Oxidative Stress Involved in Renal Ischemic-Reperfusion Injury [Pubmed:29550818]
American Journal of Translational?Research Advanced maternal age impairs spatial learning capacity in young adult mouse offspring [Pubmed:29636887]
Journal of?Diabetes?and its?Complications Clinical implication of alterations in serum Klotho levels in patients with type 2 diabetes mellitus and its associated complications [Pubmed:30042059]
Medical?Science?Monitor Klotho Inhibits Proliferation and Migration of Angiotensin II-Induced Vascular Smooth Muscle Cells (VSMCs) by Modulating NF-κB p65, Akt, and Extracellular Signal?… [Pubmed:30004089]
Nephrology Dialysis?Transplantation High-serum phosphate and parathyroid hormone distinctly regulate bone loss and vascular calcification in experimental chronic kidney disease [Pubmed: 30189026]
BMC?Nephrology The role of klotho in chronic kidney disease [Pubmed: 30348110]
Nature?Communications Age-related declines in α-Klotho drive progenitor cell mitochondrial dysfunction and impaired muscle regeneration [Pubmed: 30451844]
Kidney?International Wnt/β-catenin signaling mediates both heart and kidney injury in type 2 cardiorenal syndrome [Pubmed: 30770217]
Expert?Opinion on?Therapeutic?Targets The use of LP533401 as a therapeutic option for renal osteodystrophy affects, renal calcium handling, vitamin D metabolism and bone health in uremic rats [Pubmed: 30801205]
Mol Immunol CD317 mediates immunocytolysis resistance by RICH2/cytoskeleton-dependent membrane protection [Pubmed: 33223223]
Journal of Clinical Hypertension Effects of salt intervention on serum levels of Klotho influenced by salt sensitivity [Pubmed: 33164306]
CURRENT EYE RESEARCH Expression of α-Klotho is downregulated and associated with oxidative stress in the lens in streptozotocin-induced diabetic rats [Pubmed: 32744464]
international immunopharmacology Klotho ameliorates the onset and progression of cataract via suppressing oxidative stress and inflammation in the lens in streptozotocin-induced diabetic rats [Pubmed: 32438076]
Purinergic Signalling P2X7 siRNA targeted to the kidneys increases klotho and delays the progression of experimental diabetic nephropathy. [Pubmed: 32377919]
Oxidative Medicine and Cellular Longevity Ameliorating Effect of Klotho Protein on Rat Heart during I/R Injury [Pubmed: 33123314]
NEUROSCIENCE Increased Plasma Level of Longevity Protein Klotho as a Potential Indicator of Cognitive Function Preservation in Patients With Schizophrenia [Pubmed: 32612508]
Pharmacological Reports |áKlotho protein has therapeutic activity in contrast-induced acute kidney injury by limiting NLRP3 inflammasome-mediated pyroptosis and promoting autophagy [33675964]
J Mol Neurosci Investigation of the Role of Induced Overexpression of the Isolated Secreted Klotho on the A-172 Human Glioblastoma Cells [Pubmed:35112226]
Reproductive Biology and Endocrinology Expression of Klotho in Follicles of Patients With Polycystic Ovary Syndrome and Its Clinical Significance []
Journal of Ovarian Research Klotho is a prognostic indicator of oocytes qualities in polycystic ovary syndrome []
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