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中英文说明书丨艾美捷FD Apop TUNEL凋亡检测试剂盒
FD Apop™ Kit is designed for the microscopic detection of cells undergoing apoptosis based on the principle of in situ DNA nick-end labeling (TUNEL) technique¹. The assay uses terminal deoxynucleotidyl transferase to catalyze the incorporation of biotinylated deoxyuridines onto the free 3′-hydroxyl termini of DNA fragments, which are considered one of the most characteristic features of apoptosis2, 3. The integrated biotins are amplified and visualized with the avidin-biotin-complex (ABC) method4, enabling light microscopic identification.
The reagents and procedure of FD Apop™ Kit have been optimized to achieve a high degree of both specificity and sensitivity for detecting apoptotic cells with minimal background. This kit can be used with frozen and paraffin sections, as well as cultured cells. The procedure of the kit takes approximately 4 hours.
Kit contents:
Part I (Store at -20°C)
Digestive Enzyme 2 ml x 4
Reaction Solution A 2 ml x 2
Reaction Solution B 60 µl
Reaction Solution C 40 µl
Chromogen Solution 20 ml
Part II (Store at 4°C)
Equilibration Buffer 20 ml
Detection Reagent 6 ml
10x Phosphate-Buffered Saline 250 ml x 2
艾美捷FD NeuroTechFD Apop TUNEL凋亡检测试剂盒(#PK101)设计用于基于原位DNA切口标记(TUNEL)技术原理的细胞凋亡的显微镜检测。该测定使用末端脱氧核苷酸转移酶催化生物素化脱氧尿苷掺入DNA片段的游离3′-羟基末端,这被认为是细胞凋亡极为典型的特征之一2, 3.整合的生物素通过亲和素-生物素复合物(ABC)方法进行扩增和可视化4,可实现光学显微镜识别。
FD NeuroTech FD Apop TUNEL凋亡检测试剂盒的试剂和程序已经过优化,可实现高度的特异性和灵敏度,以极小的背景检测凋亡细胞。该试剂盒可用于冷冻和石蜡切片以及培养细胞。该套件的过程大约需要 4 小时。
FD Apop TUNEL凋亡检测试剂盒文献引用:
1.Gavrieli Y, Sherman Y, and Ben-Sasson SA. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119:493-501.
2.Wyllie AH. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 284:555-6.
3.Arends MJ, Morris RG, and Wyllie AH. (1990) Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136:593-608.
4.Hsu SM, Raine L, and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29:577-80.
来源:https://www.amyjet.com/products/FDN-PK101.shtml
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中英文说明书丨艾美捷FD NeuroApap神经元TUNEL凋亡检测试剂盒
FD NeuroApop™ Kit is specifically designed for the detection of neuronal apoptosis in tissue sections from the central nervous system based on the principle of in situ DNA nick-end labeling (TUNEL) technique. The assay uses terminal deoxynucleotidyl transferase to catalyze the incorporation of biotinylated deoxyuridines onto the free 3′-hydroxyl termini of DNA fragments, which are considered one of the most characteristic features of apoptosis. The integrated biotins are amplified and visualized with the avidin-biotin-complex (ABC) method4, enabling light microscopic identification.
The reagents and procedure of FD NeuroApop™ Kit have been optimized to achieve a high degree of both specificity and sensitivity for detecting apoptotic neurons with the lowest background. This kit can be used with frozen and paraffin sections, as well as cultured cells (cf. photo samples below). The procedure of the kit takes approximately 4 hours.
Kit contents:
Part I (Store at -20°C)
Digestive Enzyme 2 ml x 4
Reaction Solution A 2 ml x 2
Reaction Solution B 85 µl
Reaction Solution C 60 µl
Chromogen Solution 20 ml
Part II (Store at 4°C)
Equilibration Buffer 20 ml
Detection Reagent 5 ml
10x Phosphate-Buffered Saline 250 ml x 2
艾美捷FD NeuroTech FD NeuroApap 神经元TUNEL凋亡检测试剂盒(#PK201)专为检测中枢神经系统组织切片中的神经元凋亡而设计,基于原位DNA切口末端标记(TUNEL)技术的原理。该测定使用末端脱氧核苷酸转移酶催化生物素化脱氧尿苷掺入DNA片段的游离3′-羟基末端,这被认为是细胞凋亡比较典型的特征之一 2, 3.整合的生物素通过亲和素-生物素复合物(ABC)方法进行扩增和可视化4,可实现光学显微镜识别。
FD NeuroApap 神经元TUNEL凋亡检测试剂盒(#PK201)的试剂和程序已经过优化,在检测具有极低背景的凋亡神经元时实现了高度的特异性和灵敏度。该试剂盒可用于冷冻和石蜡切片以及培养细胞(参见下面的照片样品)。该套件的过程大约需要 4 小时。
FD NeuroApap 神经元TUNEL凋亡检测试剂盒文献引用:
1.Gavrieli Y, Sherman Y, and Ben-Sasson SA. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119:493-501.
2.Wyllie AH. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 284:555-6.
3.Arends MJ, Morris RG, and Wyllie AH. (1990) Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136:593-608.
4.Hsu SM, Raine L, and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29:577-80.
来源:https://www.amyjet.com/products/FDN-PK201.shtml
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中英文说明书丨艾美捷FD NeuroSilver退化神经元银染试剂盒II
FD NeuroSilver™ Kit II is designed for the detection of degenerating neurons in fixed tissue sections of the central nervous system from experimental animals. The principle of this kit is based on the findings that some components of the neurons undergoing degeneration, such as lysosomes, axons, and terminals, become particularly argyrophilic. Under certain conditions, these cellular elements bind to the silver ions with high affinity. Upon reduction, the silver ions form metallic grains that are visible under a light or electron microscope.
FD NeuroSilver™ Kit II has been used widely in animal studies under various experimental conditions. This kit has proven extremely specific and sensitive for the detection of degenerating neuronal somata, axons, and terminals in both the brain and the spinal cord (cf. photo samples). It is particularly useful for the detection of small numbers of degenerating neurons that may not be demonstrable with routine histopathological techniques.
FD NeuroSilver™ Kit II has also been proven to be very sensitive and reliable for the detection of amyloid plaques in the brain of transgenic mice (cf. photo samples). In addition, this kit may be used for demonstrating neurodegeneration and/or amyloid plaques in tissue sections that have been processed for immunohistochemistry (cf. photo samples). The procedure of FD NeuroSilver™ Kit II takes approximately 1 hour.
Kit contents:
Solution A 500 ml
Solution B 500 ml
Solution C 500 ml x 2
Solution D 500 ml
Solution E 2 ml
Solution F 3 ml
Solution G (10X) 500 ml
Glass Specimen Retriever 2
Natural hair paintbrush 1
User Manual 1
艾美捷FD NeuroTech FD NeuroSilver退化神经元银染试剂盒II(#PK301)设计用于检测实验动物中枢神经系统固定组织切片中的退化神经元。该试剂盒的原理是基于以下发现:经历退化的神经元的某些成分,如溶酶体、轴突和末端,变得特别嗜银。在某些条件下,这些细胞元素以高亲和力与银离子结合。还原后,银离子形成金属颗粒,在光学或电子显微镜下可见。
FD NeuroSilver退化神经元银染试剂盒II已广泛用于各种实验条件下的动物研究。该试剂盒已被证明对检测大脑和脊髓中的退化神经元体瘤、轴突和末端具有极高的特异性和灵敏度。它对于检测少量退化神经元特别有用,这些神经元可能无法通过常规组织病理学技术证明。
FD NeuroSilver退化神经元银染试剂盒II也被证明对检测转基因小鼠大脑中的淀粉样斑块非常敏感和可靠(参见照片样品)。此外,该试剂盒可用于显示已处理用于免疫组织化学的组织切片中的神经变性和/或淀粉样斑块。FD神经银™试剂盒II的过程大约需要1小时。
来源:https://www.amyjet.com/products/FDN-PK301.shtml
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中英文说明书丨艾美捷FD快速高尔基法染色试剂盒
Golgi-Cox impregnation1, 2 has been one of the most effective techniques for studying both the normal and abnormal morphology of neurons as well as glia. Using the Golgi technique, subtle morphological alterations in neuronal dendrites and dendritic spines have been discovered in the brains of animals treated with drugs as well as in the postmortem brains of patients with neurological diseases3, 4. However, the unreliability and the time-consuming process of Golgi staining have been major obstacles to the widespread application of this technique.
FD Rapid GolgiStain™ Kit is designed based on the principle of the methods described by Ramón- Moliner2, Glaser and Van der Loos5. This kit has not only dramatically improved and simplified the Golgi-Cox technique but has also proven to be extremely reliable and sensitive for demonstrating morphological details of neurons and glia, especially dendritic spines. The FD Rapid GolgiStain™ Kit has been tested extensively and widely used on the brains from several species of animals as well as on the specimens of postmortem human brains.
Kit contents:
Store at room temperature
Solution A 250 ml
Solution B 250 ml
Solution C 250 ml x 2
Solution D 250 ml
Solution E 250 ml
Glass Specimen Retriever 2
Natural hair paintbrush 2
Dropping bottle 1
User Manual 1
艾美捷FD NeuroTech FD快速高尔基法染色试剂盒(#PK401)背景:
高尔基体-考克斯浸渍1, 2一直是研究神经元以及神经胶质细胞正常和异常形态的有效技术之一。使用高尔基体技术,在接受药物治疗的动物大脑以及神经系统疾病患者的死后大脑中发现了神经元树突和树突棘的细微形态变化3, 4.然而,高尔基体染色的不可靠性和耗时的过程一直是该技术广泛应用的主要障碍。
FD快速高尔基法染色试剂盒是根据Ramón-Moliner描述的方法原理设计的2、格拉泽和范德洛斯5.该试剂盒不仅显着改进和简化了高尔基体-考克斯技术,而且还被证明在展示神经元和神经胶质细胞(尤其是树突棘)的形态细节方面非常可靠和灵敏。FD 快速高尔基体染色™试剂盒已在多种动物的大脑以及死后人类大脑标本上进行了广泛和广泛的测试。
来源:https://www.amyjet.com/products/FDN-PK401.shtml
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艾美捷Detroit R&D氧化应激ELISA(8-异丙肾上腺素)试剂盒
艾美捷Detroit R&D氧化应激ELISA(8-异丙肾上腺素)试剂盒(##8iso1)用于测定生物样品中的8-异丙肾上腺素水平。典型的标准曲线酶联免疫吸附试验的结果如下:
异丙烷类是由组织的随机氧化产生的非酶源类二十烷类由氧自由基产生的磷脂 前列腺素F2a是体内氧化损伤和相关的指标冠状动脉钙化(CAC)增加。最近一项使用8-异丙肾上腺素酶联免疫吸附法的研究表明,高脂肪大鼠肝脏8-异丙烷水平增加(HF)饮食诱导的非酒精性脂肪性肝病(NAFLD)。
有趣的是,血管紧张素II受体替米沙坦拮抗剂(高血压药物)显著降低肝脏8-异丙肾上腺素水平。8的尿液排泄-醛固酮输注诱导的小鼠肾纤维化导致异丙肾上腺素增加。8-异丙肾上腺素排泄水平在骨桥蛋白(参与肿瘤促进)敲除小鼠中显著降低,这表明骨桥蛋白是一种氧化应激诱导的肾损伤的治疗靶点 肾脏NADPH氧化酶亚单位mRNA的水平链脲佐菌素诱导的糖尿病(DM)中8-异丙肾上腺素的表达和尿8-异丙斯坦的增加. 8-异丙肾上腺素已被检测为运动诱导的肌肉损伤生物标志物和污水生物标志物用于评估社区健康Yamamato等人表明AST-120的使用与接受维持性血液透析治疗的患者的氧化应激(8-异丙肾上腺素水平下降)。
高脂血症小鼠的子宫内柴油废气暴露不会影响后期的氧化还原稳态。一个强壮的德国一项针对老年人的大型研究发现,吸烟与尿8-异丙肾上腺素水平之间存在关联成年吸烟者。喂食高脂肪饮食的T2D诱导大鼠的比目鱼肌表现出更高水平的8-异丙肾上腺素与增加的mt-DNA拷贝数和8-OHdG6结合高氧血症增加了8-异丙肾上腺素在野生型小鼠的肺中,但不在可溶性环氧化物水解酶敲除小鼠中. 稀释的尿液或没有提取的细胞培养基适用于ELISA。
每个试剂盒适合一式三份对多达24个样品的分析包括一个96孔板、一小瓶8-异丙肾上腺素标准品、一小瓶8-异丙肾上腺素缀合的辣根过氧化物酶(HRP),用于样品和HRP稀释液的缓冲液,以及平板洗涤。
Detroit R&D氧化应激ELISA(8-异丙肾上腺素)试剂盒部分文献:
1. Gào X, Xuan Y, Benner A, Anusruti A, Brenner H,Schöttker, B. Nitric Oxide Metabolites and Lung Cancer Incidence: A Matched Case-Control Study Nested in the ESTHER Cohort. Oxidative Medicine and Cellular Longevity Volume 2019, Article ID 6470950,
2. Kawasaki K, Kondoh E, Chigusa Y, Kawamura Y, Mogami H, Takeda S, Horie A, Baba T, Matsumura N, Mandai M, Konishi I. Metabolomic profiles of placenta in preeclampsia: Antioxidant effect of magnesium sulfate on trophoblasts in early-onset preeclampsia. Hypertension 73: 671-679. 2019.
3. Gào X, Zhang Y, Burwinkel B, Xuan Y, Holleczek B, Brenner H, Schöttker B. The associations of DNA methylation alterations in oxidative stress-related genes with cancer incidence and mortality outcomes: a population-based cohort study. Clinical Epigenetics 11: 14-22. 2019.
4. Gao, X, Gào X, Zhang Y, Holleczek B, Schöttker B, Brenner H. Oxidative stress and epigenetic mortality risk score: associations with all-cause mortality among elderly people. Eur. J. Epidemiology 34: 451-462.
Detroit R&D专注于心血管疾病、癌症的研究,诊断和治疗领域。涉及环境健康科学、高血压、乳腺癌和前列腺癌相关研究领域。艾美捷科技是Detroit R&D的中国代理商,为科研工作者提供优质的产品与服务。
来源:https://www.amyjet.com/news/Detroit-R-D-new.shtml
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艾美捷Detroit R&D DNA损伤分析试剂盒解决方案
Detroit R&D DNA损伤分析试剂盒为基础研究和工业应用提供了强大的工具。这种创新的方法是基于在一个长(8.2-8.8kb)的片段上进行30个循环的定量PCR随后使用实时PCR进行定量。
艾美捷Detroit R&DDNA损伤分析试剂盒可以用于人、大鼠或小鼠的血液、细胞或组织(见下文)。DNA损伤由于缺失和链断裂,可以用该试剂盒检测。使用微孔板格式化的DNA分离技术,该试剂盒可以很容易地用于高通量总体安排在这方面,底特律研发分析是一种很有前途的工具,因为它快速、简单,而且只需要少量测试物质。
Detroit R&D DNA损伤分析试剂盒,用于通过以下方法在体内和体外测量受损的8.8kb线粒体DNA在QPCR分析之后用实时PCR对复制的DNA进行定量:每个试剂盒包含DNA聚合酶、QPCR引物和用于QPCR和实时PCR引物的dNTP以及8.8kb实时PCR标准(不包括用于实时PCR的SYBR Green主混合物)定量。对20个样品(45个反应)进行重复分析。
图:左图:实时PCR标准的扩增图;右图:8.8kb线粒体DNA实时PCR分析的标准曲线
此外,#MCN1、MCN2、MCN3线粒体DNA拷贝数试剂盒这些是基于定量PCR的试剂盒,用于测量人、大鼠或小鼠的线粒体DNA拷贝数小鼠该试剂盒包括产生200kb线粒体DNA的小鼠、人或大鼠引物序列此外,该试剂盒包括核DNA(B-肌动蛋白)序列的引物,内部控制对于标准曲线,含有Taq DNA聚合酶、dNTP、MgCl2和缓冲液的反应混合物,以及化验板。
Detroit R&D DNA损伤分析试剂盒相关研究:
#DD2H:人类QPCR后的实时PCR DNA定量
#DD2M:小鼠QPCR后的实时PCR DNA定量
#DD2R:大鼠QPCR后的实时PCR DNA定量
Detroit R&D专注于心血管疾病、癌症的研究,诊断和治疗领域。涉及环境健康科学、高血压、乳腺癌和前列腺癌相关研究领域。艾美捷科技是Detroit R&D的中国代理商,为科研工作者提供优质的产品与服务。
来源:https://www.amyjet.com/news/Detroit-R-D-new.shtml
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艾美捷Detroit R&D 柠檬酸盐合酶测定解决方案
位于线粒体基质的柠檬酸合成酶是线粒体中的限速酶柠檬酸循环(或克雷布斯循环)的第一步。柠檬酸合成酶催化乙酰辅酶A和草酰乙酸的乙酸残基形成柠檬酸盐。醋酸氧苄酯在完成一轮克雷布斯循环后再生。酶活性用于评估线粒体的氧化能力以及线粒体的完整性。
柠檬酸合成酶可以通过以下方式测量从CoA-SH释放的-SH基团的形成使用反应性埃尔曼试剂(5,5'-二硫代双[2-硝基苯甲酸],DTNB)并监测吸光度在412纳米处。
艾美捷Detroit R&D 柠檬酸盐合酶测定组分如下:
•96孔板。
•提取缓冲液。
•反应缓冲液。
•DTNB混合。
•乙酰辅酶A
•氧卤乙酸盐混合物
样品制备:
•硬组织:在刀式匀浆器(Polytron、Turrax或快速超声处理)中以每克组织4毫升提取缓冲液的比例,并在4°C下以1000克离心15分钟。收集上清液,测量蛋白质(最佳浓度约为0.5 mg/ml)。
•细胞:在Potter Elvehjem中超声或均质。在4°C下以1000g离心10分钟,收集上清液,测量蛋白质(准确浓度约为0.5mg/ml)。
•富含线粒体的部分:由于92%的柠檬酸合成酶活性位于线粒体和乙酰辅酶A不能穿透其中,可以通过分离线粒体来制备(提高产量)并将其分解。用线粒体匀浆对样品进行匀浆缓冲液(250mM蔗糖、2mM乙二胺四乙酸(EDTA)和25mM Tris-HCl,pH 7.4)加蛋白酶抑制剂。在4ºC下以800g离心10分钟。仔细收集上清液(丢弃颗粒),并在4ºC下以15.000-20.000 x g离心20分钟。通过添加清洗颗粒PBS,离心15.000-20.000 x g,离心5分钟。用提取缓冲液重新悬浮颗粒。
试剂制备:
•解冻所有试剂。
•在DTNB混合管中加入985µl反应缓冲液(在冰上稳定一天)。
含量测定(415 nm,37℃):
• 20ml样品(总蛋白质范围5-20µg/ml)
•10µl反应缓冲液
• 5 m乙酰辅酶A的l
• 15ml样品缓冲液
•增加100ml至板材坯料
•混合10秒,在37ºC下孵育1分钟。
•增加50ml草酰乙酸盐混合物,快速混合5秒,在3-8分钟内读取。
反应空白-添加提取缓冲液代替反应缓冲液中的样品以及乙酰辅酶A和草酰乙酸混合样品(除ELISA板空白外)应从样品中扣除该活性价值观
Detroit R&D 柠檬酸盐合酶测定文献引用:
[1] PA Srere.Citrate synthase.Methods Enzymol., 13:3-11, 1969.
[2] D Shepherd & PB Garland.Citrate synthase from rat liver. Methods Enzymol., 13:11-16, 1969.
[3] E Bogin & A Wallace.Citrate synthase from lemon fruit. Methods Enzymol., 13:19-22, 1969.
[4] J. Santos et al. Free Radical Biology and Medicine 51: 1849-1860, 2011
Detroit R&D专注于心血管疾病、癌症的研究,诊断和治疗领域。涉及环境健康科学、高血压、乳腺癌和前列腺癌相关研究领域。艾美捷科技是Detroit R&D的中国代理商,为科研工作者提供优质的产品与服务。
来源:https://www.amyjet.com/news/Detroit-R-D-new.shtml
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艾美捷Detroit R&D GLOBRITE ECL试剂盒解决方案
艾美捷Detroit R&D GLOBrite化学发光试剂盒结合了高灵敏度 成本低,使用方便。该 ecl 试剂盒用于检测固定化抗原 与辣根过氧化物酶标记的抗体偶联。只需 1:1 混合每种 ECL 试剂 使用前。
Detroit R&D GLOBRITE ECL试剂盒易于使用:
•简单地混合等量的试剂1和2
•直接应用于薄膜
Detroit R&D 研发化学发光的比较,试剂盒与竞争对手试剂盒的对比,两分钟的曝光时间
保存:
如果所有组件在使用前都储存在2至8摄氏度的适当温度下,该套件将获得极佳效果。
收到此试剂盒后,应将物品存放在指定的温度下。
提供的材料:
项目 描述 数量
检测试剂1 50 mL(GLB1)或125 mL(GLB2)2
检测试剂2 50 mL(GLB1)或125 mL(GLB2)2
注意事项:
1.在开始分析之前,请仔细阅读所有说明。
2.该试剂盒中的试剂经过测试和配制,以达到最佳效果。此套件可能不起作用
3.如果更换了任何试剂或修改了任何程序,则正确。
4.本试剂盒仅供研究使用,不得用于诊断。
1) 应准备足以覆盖膜的ECL试剂量。
2) 配制后立即使用混合试剂。
3) 混合等量的试剂1和试剂2
4) 在室温下将膜在ECL试剂中孵育5分钟。
5) 移除膜并从膜中排出多余的ECL试剂
6) 将薄膜放在塑料片中(蛋白质面朝上)。
7) 如果使用胶卷,请将胶卷暴露在暗室中,然后放入胶卷暗盒中
8) 在发育之前培养适当的时间。最佳孵育时间取决于所使用的第一抗体和第二抗体。建议第一次接触时间为1分钟。然后可以基于该初始曝光来调整曝光时间。
9) 如果使用胶片成像仪,请按照步骤5将胶片放置在成像仪内,并曝光适当的时间长度。
Detroit R&D GLOBRITE ECL试剂盒文献引用:
Towbin, H. and Gordon, J. (1984). Immunoblotting and dot immunoblotting – Current status and outlook. J.Immunol. Meth. 72:313-40.
Young, P.R. (1989) An improved method for the detection of peroxidase conjugated antibodies on immunoblots. J. Virol. Meth. 24:227-36.
Detroit R&D专注于心血管疾病、癌症的研究,诊断和治疗领域。涉及环境健康科学、高血压、乳腺癌和前列腺癌相关研究领域。艾美捷科技是Detroit R&D的中国代理商,为科研工作者提供优质的产品与服务。
来源:https://www.amyjet.com/news/Detroit-R-D-new.shtml