绵羊（Sheep）5核苷酸酶（5-NT）-NEWA

绵羊（Sheep）5核苷酸酶（5-NT）-NEWA 3. 细胞上清液,3000转离心10分钟去除颗粒和聚合物。 本试剂盒只能用于科学研究,不得用于医学诊断 4. 组织匀浆,将组织加入适量生理盐水捣碎。3000转离心10分钟绵羊，Sheep，5核苷酸酶，5-NT，ELISA检测试剂盒 使用说明 书 关于书的成语关于读书的排比句社区图书漂流公约怎么写关于读书的小报汉书pdf 取上清。 检测原理 5. 保存,如果样本收集后不及时检测,请按一次用量分装,冻存于 试剂盒采用双抗体一步夹心法酶联免疫吸附试验，ELISA，。往预-20?,避免反复冻融,在室温下解冻并确保样品均匀地充分解冻。 先包被5核苷酸酶，5-NT，抗体的包被微孔中,依次加入标本、 标准 excel标准偏差excel标准偏差函数exl标准差函数国标检验抽样标准表免费下载红头文件格式标准下载 自备物品 品、HRP标记的检测抗体,经过温育并彻底洗涤。用底物TMB显色,1. 酶标仪，450nm， 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2. 高精度加样器及枪头,0.5-10uL、2-20uL、20-200uL、200-1000uL 的黄色。颜色的深浅和样品中的5核苷酸酶，5-NT，呈正相关。用酶3. 37?恒温箱 标仪在450nm 波长下测定吸光度，OD 值，,计算样品浓度。 操作注意事项 样品收集、处理及保存 方法 快递客服问题件处理详细方法山木方法pdf计算方法pdf华与华方法下载八字理论方法下载 1. 试剂盒保存在2-8?,使用前室温平衡20分钟。从冰箱取出的1. 血清,使用不含热原和内毒素的试管,操作过程中避免任何细胞浓缩洗涤液会有结晶,这属于正常现象,水浴加热使结晶完全溶解刺激,收集血液后,3000转离心10分钟将血清和红细胞迅速小心地后再使用。 分离。 2. 实验中不用的板条应立即放回自封袋中,密封，低温干燥，保存。 2. 血浆,EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。 3. 浓度为0的S0号标准品即可视为阴性对照或者空白,按照说明 书操作时样本已经稀释5倍,最终结果乘以5才是样本实际浓度。 1. 手工洗板,甩尽孔内液体,每孔加满洗涤液,静置1min后甩尽4. 严栺按照说明书中标明的时间、加液量及顺序进行温育操作。 孔内液体,在吸水纸上拍干,如此洗板5次。 5. 所有液体组分使用前充分摇匀。 2. 自动洗板机,每孔注入洗液350μL,浸泡1min,洗板5次。 试剂盒组成 操作步骤 1. 从室温平衡20min后的铝箔袋中取出所需板条,剩余板条用自封名称 96孔配置 48孔配置 备注 微孔酶标板 12孔×8条 12孔×4条 无 袋密封放回4?。 标准品 0.3mL*6管 0.3mL*6管 无 样本稀释液 6mL 3mL 无 2. 设置标准品孔和样本孔,标准品孔各加不同浓度的标准品50μL, 检测抗体-HRP 10mL 5mL 无 20×洗涤缓冲液 25mL 15mL 按说明书进行稀释 3. 样本孔先加待测样本10μL,再加样本稀释液40μL,空白孔不底物A 6mL 3mL 无 底物B 6mL 3mL 无 加。 终止液 6mL 3mL 无 封板膜 2张 2张 无 4. 除空白孔外,标准品孔和样本孔中每孔加入辣根过氧化物酶说明书 1份 1份 无 自封袋 1个 1个 无 ，HRP，标记的检测抗体100μL,用封板膜封住反应孔,37?水浴注,标准品，S0-S5，浓度依次为,0、5、10、20、40、80 pg/mL 锅或恒温箱温育60min。 试剂的准备 5. 弃去液体,吸水纸上拍干,每孔加满洗涤液,静置1min,甩去 20×洗涤缓冲液的稀释,蒸馏水按1,20稀释,即1份的20×洗涤缓 洗涤液,吸水纸上拍干,如此重复洗板5次，也可用洗板机洗板，。 冲液加19份的蒸馏水。 6. 每孔加入底物A、B各50μL,37?避光孵育15min。 洗板方法 7. 每孔加入终止液50μL,15min内,在450nm波长处测定各孔的试剂盒性能 OD值。 1. 准确性,标准品线性回归与预期浓度相关系数R值,大于等于结果判断 0.9900。 绘制标准曲线,在Excel工作 表 关于同志近三年现实表现材料材料类招标技术评分表图表与交易pdf视力表打印pdf用图表说话 pdf 中,以标准品浓度作横坐标,对应2. 灵敏度,最低检测浓度小于1.0 pg/mL。 OD值作纵坐标,绘制出标准品线性回归曲线,按曲线方程计算各样3. 特异性,不与其它可溶性结构类似物交叉反应。 本浓度值。 4. 重复性,板内、板间变异系数均小于15%。 5. 贮藏,2-8?,避光防潮保存。 6. 有效期,6个月 免责声明 1. 试剂盒仅供研究使用,不得用于临床实验或人体实验,否则所 产生的一切后果,由实验者承担,本公司概不负责。 2. 严栺按照说明书操作,实验者违反说明书操作,后果由实验者 承担。 FOR RESEARCH USE ONLY. Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge NOT FOR USE IN DIAGNOSTIC PROCEDURES. samples for 30 minutes at 3000×g at 2-8? within 30 minutes of collection. Store samples at -20?or -80?. Avoid repeated freeze-thaw cycles. Sheep 5-Nucleotidase (5-NT) ELISA Kit instruction Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20?or Intended use -80?. Avoid repeated freeze-thaw cycles. This 5-NT ELISA kit is intended Laboratory for Research use only and is not for Note: The samples shoule be centrifugated dequately and no hemolysis use in diagnostic or therapeutic procedures.The Stop Solution changes the color or granule was allowed. from blue to yellow and the intensity of the color is measured at 450 nm using a Materials required but not supplied spectrophotometer. In order to measure the concentration of 5-NT in the sample, 1. Standard microplate reader(450nm) this 5-NT ELISA Kit includes a set of calibration standards. The calibration 2. Precision pipettes and Disposable pipette tips. standards are assayed at the same time as the samples and allow the operator to 3. 37 ? incubator produce a standard curve of Optical Density versus 5-NT concentration. The Precautions concentration of 5-NT in the samples is then determined by comparing the O.D. of 1. Do not substitute reagents from one kit to another. Standard, conjugate and the samples to the standard curve. microplates are matched for optimal performance. Use only the reagents supplied by Sample collection and storages manufacturer. Serum - Use a serum separator tube and allow samples to clot for 30 minutes 2. Do not remove microplate from the storage bag until needed. Unused strips before centrifugation for 10 minutes at approximately 3000×g. Remove serum and should be stored at 2-8?C in their pouch with the desiccant provided. assay immediately or aliquot and store samples at -20? or -80?.Avoid repeated 3. Mix all reagents before using. freeze-thaw cycles Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25?C) sample well; Blank well doesn’t add anyting. 4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip Materials supplied and incubate for 60 minutes at 37?C. Name 96 determinations 48 determinations Microelisa stripplate 12*8strips 12*4strips 5. Aspirate each well and wash, repeating the process four times for a total of five Standard 0.3ml*6tubes 0.3ml*6tubes washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, Sample Diluent 6.0ml 3.0ml manifold dispenser or autowasher. Complete removal of liquid at each step is HRP-Conjugate reagent 10.0ml 5.0ml 20X Wash solution 25ml 15ml essential to good performance. After the last wash, remove any remaining Wash Chromogen Solution A 6.0ml 3.0ml Solution by aspirating or decanting. Invert the plate and blot it against clean paper Chromogen Solution B 6.0ml 3.0ml towels. Stop Solution 6.0ml 3.0ml 6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Closure plate membrane 2 2 User manual 1 1 Gently mix and incubate for 15 minutes at 37?C. Protect from light. Sealed bags 1 1 7. Add 50μl Stop Solution to each well. The color in the wells should change Note: Standard (S0 ? S5) concentration was followed by 0,5,10,20,40,80 pg/ml. from blue to yellow. If the color in the wells is green or the color change does not Reagent preparation appear uniform, gently tap the plate to ensure thorough mixing. 20×wash solution:Dilute with Distilled or deionized water 1:20. 8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader Assay procedure within 15 minutes. 1. Prepare all reagents before starting assay procedure. It is recommended that Calculation of results all Standards and Samples be added in duplicate to the Microelisa Stripplate. 1. This standard curve is used to determine the amount in an unknown sample. 2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to The standard curve is generated by plotting the average O.D. (450 nm) standard well. obtained for each of the six standard concentrations on the vertical (Y) axis 3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing versus the corresponding concentration on the horizontal (X) axis. 2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software. 3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration. 4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve. 5. The sensitivity by this assay is 1.0 pg/ml Storage: 2-8?. 6. Standard curve validity: six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!