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補(bǔ)體成分5a(C5a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Complement Component 5a (C5a)

  • 補(bǔ)體成分5a(C5a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 補(bǔ)體成分5a(C5a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 補(bǔ)體成分5a(C5a)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
  • SEA388Hu.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)補(bǔ)體成分5a(C5a),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補(bǔ)體成分5a(C5a)(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) 90-101 95
EDTA plasma(n=5) 93-101 98
heparin plasma(n=5) 98-105 102

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)加入適量的補(bǔ)體成分5a(C5a),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補(bǔ)體成分5a(C5a)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) 95-103% 78-89% 95-102% 84-94%
EDTA plasma(n=5) 94-102% 85-98% 86-95% 92-99%
heparin plasma(n=5) 89-97% 92-101% 86-103% 92-101%

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

將補(bǔ)體成分5a(C5a)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,其中的補(bǔ)體成分5a(C5a)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的補(bǔ)體成分5a(C5a)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補(bǔ)體成分5a(C5a)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:Homo sapiens (Human,人) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
RPA388Hu01 補(bǔ)體成分5a(C5a)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
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APA388Hu61 補(bǔ)體成分5a(C5a)活性蛋白 Cell?culture;?Activity?Assays.
EPA388Hu61 補(bǔ)體成分5a(C5a)真核蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAA388Hu06 補(bǔ)體成分5a(C5a)多克隆抗體 WB; IHC; ICC; IP.
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MAA388Hu21 補(bǔ)體成分5a(C5a)單克隆抗體 WB
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MAA388Hu23 補(bǔ)體成分5a(C5a)單克隆抗體 WB; IHC; ICC; IP.
MAA388Hu24 補(bǔ)體成分5a(C5a)單克隆抗體 WB; IHC; ICC; IP.
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SEA388Hu 補(bǔ)體成分5a(C5a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA388Hu 補(bǔ)體成分5a(C5a)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.
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參考文獻(xiàn)

雜志 參考文獻(xiàn)
Molecular Immunology Cathepsin D is released after severe tissue trauma in vivo and is capable of generating C5a in vitro [ScienceDirect: S0161589011008297]
Journal of Reproductive Immunology Elevated complement factor C5a in maternal and umbilical cord plasma in preeclampsia [PubMed: 23415845]
The Journal of Clinical Investigation Complement component 5 contributes to poor disease outcome in humans and mice with pneumococcal meningitis [PubMed: PMC3195471]
Journal of Antimicrobial Chemotherapy Efficacy of a synthetic antimicrobial peptidomimetic versus vancomycin in a Staphylococcus epidermidis device-related murine peritonitis model [Pubmed: 23645587]
Journal of Orthopaedic Research Conversion from external fixator to intramedullary nail causes a second hit and impairs fracture healing in a severe trauma model [Pubmed: 23070742]
SpringerPlus Therapeutic effects of sesame oil on monosodium urate crystal-induced acute inflammatory response in rats [Pubmed: 24353977]
American Journal fo Reproductive Immunology Evaluation of the Markers of Inflammation in the Umbilical Cord Blood of Newborns of Mothers with Thrombophilia [Pubmed:25209155]
Gynecol Obstet Invest Complement Split Products C3a/C5a and Receptors: Are They Regulated by Circulating Angiotensin II Type 1 Receptor Autoantibody in Severe Preeclampsia? [PubMed: 26485247]
Journal of Neuroimmunology Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitis [Pubmed:27235358]
International Immunopharmacology Consistency and pathophysiological characterization of a rat polymicrobial sepsis model via the improved cecal ligation and puncture surgery [Pubmed:26802602]
Digital Repository Dosagem de fra??es ativadas do sistema complemento em empiema induzido em ratos [10183]
Environmental Pollution Cadmium-induced immune abnormality is a key pathogenic event in human and rat models of preeclampsia [pubmed:27511439]
Cancer Letters Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expression [pubmed:29031586]
Acta Paediatrica Serum complement factor 5a levels are associated with nonalcoholic fatty liver disease in obese children [DOI:?10.1111/apa.14106]
Scientific Reports Splenectomy modulates early immuno-inflammatory responses to trauma-hemorrhage and protects mice against secondary sepsis [Pubmed: 30291296]
Nutrients The Beneficial Effects of Lactobacillus plantarum PS128 on High-Intensity, Exercise-Induced Oxidative Stress, Inflammation, and Performance in Triathletes [Pubmed: 30736479]
Journal of?NeuroVirology Intrathecal complement activation by the classical pathway in tick-borne encephalitis [Pubmed: 30850976]
PLoS One Sepsis causes right ventricular myocardial inflammation independent of pulmonary hypertension in a porcine sepsis model [Pubmed: 31247004]
Frontiers in Immunology C5a/C5aR Pathway is Critical for the Pathogenesis of Psoriasis [Pubmed: 31447855]
Phytomedicine Novel evidence that an alternative complement cascade pathway is involved in optimal mobilization of hematopoietic stem/progenitor cells in Nlrp3?… []
Aging C5aR deficiency attenuates the breast cancer development via the p38/p21 axis [Pubmed: 32669478]
STEM CELL REVIEWS AND REPORTS A Novel Evidence That Mannan Binding Lectin (MBL) Pathway of Complement Cascade Activation is Involved in Homing and Engraftment of Hematopoietic Stem … [Pubmed: 32406006]
Modulation of C5a-C5aR interactions against murine mammary cancer cell line []
FASEB J C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease [Pubmed:35503088]
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