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补体成分5a(C5a)检测试剂盒(酶联免疫吸附试验法)

ELISA Kit for Complement Component 5a (C5a)

  • 补体成分5a(C5a)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • 补体成分5a(C5a)检测试剂盒(酶联免疫吸附试验法) 产品包装(模拟)
  • 补体成分5a(C5a)检测试剂盒(酶联免疫吸附试验法) 实验结果图
  • SEA388Po.jpg 标准曲线图
  • Certificate 通过ISO 9001、ISO 13485质量体系认证

特异性

本试剂盒用于检测补体成分5a(C5a),经检测与其它相似物质无明显交叉反应。
由于受到技术及样本来源的限制,不可能完成对所有相关或相似物质交叉反应检测,因此本试剂盒有可能与未经检测的其它物质有交叉反应。

回收率

分别于定值血清及血浆样本中加入一定量的补体成分5a(C5a)(加标样品),重复测定并计算其均值,回收率为测定值与理论值的比率。

样本 回收率范围(%) 平均回收率(%)
serum(n=5) 80-97 91
EDTA plasma(n=5) 90-104 101
heparin plasma(n=5) 93-101 98

精密度

精密度用样品测定值的变异系数CV表示。CV(%) = SD/mean×100
批内差:取同批次试剂盒对低、中、高值定值样本进行定量检测,每份样本连续测定20 次,分别计算不同浓度样本的平均值及SD值。
批间差:选取3个不同批次的试剂盒分别对低、中、高值定值样本进行定量测定,每个样本使用同一试剂盒重复测定8次,分别计算不同浓度样本的平均值及SD值。
批内差: CV<10%
批间差: CV<12%

线性

在定值血清及血浆样本内加入适量的补体成分5a(C5a),并倍比稀释成1:2,1:4,1:8,1:16的待测样本,线性范围即为稀释后样本中补体成分5a(C5a)含量的测定值与理论值的比率。

样本 1:2 1:4 1:8 1:16
serum(n=5) 92-105% 81-89% 95-102% 79-98%
EDTA plasma(n=5) 87-103% 78-99% 93-101% 81-101%
heparin plasma(n=5) 78-93% 98-105% 82-95% 88-101%

稳定性

经测定,试剂盒在有效期内按推荐温度保存,其活性降低率小于5%。
为减小外部因素对试剂盒破坏前后检测值的影响,实验室的环境条件需尽量保持一致,尤其是实验室内温度、湿度及温育条件。其次由同一实验员来进行操作可减少人为误差。

实验流程

1. 实验前标准品、试剂及样本的准备;
2. 加样(标准品及样本)100µL,37°C孵育1小时;
3. 吸弃,加检测溶液A100µL,37°C孵育1小时;
4. 洗板3次;
5. 加检测溶液B100µL,37°C孵育30分钟;
6. 洗板5次;
7. 加TMB底物90µL,37°C孵育10-20分钟;
8. 加终止液50µL,立即450nm读数。

实验原理

将补体成分5a(C5a)抗体包被于96孔微孔板中,制成固相载体,向微孔中分别加入标准品或标本,其中的补体成分5a(C5a)与连接于固相载体上的抗体结合,然后加入生物素化的补体成分5a(C5a)抗体,将未结合的生物素化抗体洗净后,加入HRP标记的亲和素,再次彻底洗涤后加入TMB底物显色。TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的补体成分5a(C5a)呈正相关。用酶标仪在450nm波长下测定吸光度(O.D.值),计算样品浓度。

相关产品

编号 适用物种:Sus scrofa; Porcine (Pig,猪) 应用(仅供研究使用,不用于临床诊断!)
RPA388Po01 补体成分5a(C5a)重组蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAA388Po01 补体成分5a(C5a)多克隆抗体 WB; IHC; ICC; IP.
SEA388Po 补体成分5a(C5a)检测试剂盒(酶联免疫吸附试验法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA388Po 补体成分5a(C5a)等多因子检测试剂盒(流式荧光发光法) FLIA Kit for Antigen Detection.

参考文献

杂志 参考文献
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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 frações 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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