本试剂盒只能用于科学研究，不得用于医学诊断本试剂盒只能用于科学研究，不得用于医学诊断大豆球蛋白（大豆球蛋白（globulinglobulin））ELISAELISA 检测试剂盒检测试剂盒使用说明书检测原理检测原理试剂盒采用双抗体一步夹心法酶联免疫吸附试验（ELISA） 往预先包被大豆球蛋白（globulin）抗体的包被微孔中，依次加入标本、标准品、HRP标记的检测抗体，经过温育并彻底洗涤用底物TMB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大豆球蛋白（globulin）呈正相关用酶标仪在450nm 波长下测定吸光度（OD 值） ，计算样品浓度样品收集、处理及保存方法样品收集、处理及保存方法1. 样本不能含叠氮钠（NaN3） ，因为叠氮钠（NaN3）是辣根过氧化物酶（HRP）的抑制剂2. 标本采集后尽早进行提取，提取按相关文献进行3. 植物萃取液或其它相关样本：请1000 x g离心20分钟，取上清即可检测4. 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-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自备物品自备物品1.酶标仪（450nm）2.高精度加样器及枪头：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒温箱操作注意事项操作注意事项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后再使用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4. 严格按照说明书中标明的时间、加液量及顺序进行温育操作5. 所有液体组分使用前充分摇匀试剂盒组成试剂盒组成名称名称9696 孔配置孔配置4848 孔配置孔配置备注备注微孔酶标板12 孔×8 条12 孔×4 条无标准品0.3mL*6 管0.3mL*6 管无样本稀释液6mL3mL无检测抗体-HRP10mL5mL无20×洗涤缓冲液25mL15mL按说明书进行稀释底物 A6mL3mL无底物 B6mL3mL无终止液6mL3mL无封板膜2 张2 张无说明书1 份1 份无自封袋1 个1 个无注：标准品（S0-S5）浓度依次为：0、50、100、200、400、800 ng/mL试剂的准备试剂的准备20×洗涤缓冲液的稀释：蒸馏水按 1：20 稀释，即 1 份的 20×洗涤缓冲液加 19 份的蒸馏水洗板方法洗板方法1. 手工洗板：甩尽孔内液体，每孔加满洗涤液，静置 1min 后甩尽孔内液体，在吸水纸上拍干，如此洗板 5 次。
2. 自动洗板机：每孔注入洗液 350μL，浸泡 1min，洗板 5 次操作步骤操作步骤1. 从室温平衡 20min 后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回 4℃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；3. 样本孔先加待测样本 10μL，再加样本稀释液 40μL；空白孔不加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（HRP）标记的检测抗体 100μL，用封板膜封住反应孔，37℃水浴锅或恒温箱温育 60min5. 弃去液体，吸水纸上拍干，每孔加满洗涤液，静置 1min，甩去洗涤液，吸水纸上拍干，如此重复洗板 5 次（也可用洗板机洗板） 6. 每孔加入底物 A、B 各 50μL，37℃避光孵育 15min7. 每孔加入终止液 50μL，15min 内，在 450nm 波长处测定各孔的 OD 值结果判断结果判断绘制标准曲线：在 Excel 工作表中，以标准品浓度作横坐标，对应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本浓度值试剂盒性能试剂盒性能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0.9900。
2. 灵敏度：最低检测浓度小于 1.0 ng/mL3. 特异性：不与其它可溶性结构类似物交叉反应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. 严格按照说明书操作，实验者违反说明书操作，后果由实验者承担FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.Soybean globulin ELISA Kit instructionIntended useThis globulin ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of globulin in the sample, this globulin ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus globulin concentration. The concentration of globulin in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storages1.Can’t detect the samples which contain NaN3, because NaN3 inhibits HRP activity of the horseradish peroxidase.2. Extract as soon as possible after Specimen collection, Extracted according to the relevant literature.Cell culture supernates and plant exact fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw.Materials required but not supplied1. Standard microplate reader(450nm)2. Precision pipettes and Disposable pipette tips.3. 37 ℃ incubatorPrecautions1. Do not substitute reagents from one kit to another. Standard, conjugate and microplates are matched for optimal performance. Use only the reagents supplied by manufacturer.2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.3. Mix all reagents before using.Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)Materials suppliedName96 determinations48 determinationsMicroelisa stripplate12*8strips12*4stripsStandard0.3ml*6tubes0.3ml*6tubesSample Diluent6.0ml3.0mlHRP-Conjugate reagent10.0ml5.0ml20X Wash solution25ml15mlChromogen Solution A6.0ml3.0mlChromogen Solution B6.0ml3.0mlStop Solution6.0ml3.0mlClosure plate membrane22User manual11Sealed bags11 Note: Standard (S0 → S5) concentration was followed by:0,50,100,200,400,800 ng/mlReagent preparation20×wash solution:Dilute with Distilled or deionized water 1:20.Assay procedure1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesn’t add anyting.4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each s。