猪（Porcine）白细胞介素6（IL-6）ELISA检测豪运国际本豪运国际只能用于科学研究，不得用于医学诊断检测原理豪运国际采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被白细胞介素6（IL-6）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＊终的黄色。颜色的深浅和样品中的白细胞介素6（IL-6）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分离。2. 血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。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-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1. 豪运国际保存在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，＊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。豪运国际组成名称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 pg/mL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60min。5. 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15min。7. 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 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。 豪运国际性能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：＊低检测浓度小于1.0 pg/mL。3. 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1. 豪运国际仅供研究使用，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. Porcine Interleukin 6 (IL-6) ELISA Kit instruction Intended useThis IL-6 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 IL-6 in the sample, this IL-6 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 IL-6 concentration. The concentration of IL-6 in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge 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.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.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 bags11Note: Standard (S0 → S5) concentration was followed by:0,50,100,200,400,800 pg/ml.Reagent 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 step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis 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/ml6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
人（Human）胶原蛋白三螺旋重复包含蛋白1（CTHRC1）ELISA检测豪运国际本豪运国际只能用于科学研究，不得用于医学诊断检测原理豪运国际采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被胶原蛋白三螺旋重复包含蛋白1（CTHRC1）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＊终的黄色。颜色的深浅和样品中的胶原蛋白三螺旋重复包含蛋白1（CTHRC1）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分离。2. 血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。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-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1. 豪运国际保存在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，＊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。豪运国际组成名称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、25、50、100、200、400 ng/mL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60min。5. 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15min。7. 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 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。 豪运国际性能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：＊低检测浓度小于5.0 ng/mL。3. 检测范围：5-400 ng/mL。4. 特异性：不与其它可溶性结构类似物交叉反应。5. 重复性：板内、板间变异系数均小于15%。6. 贮藏：2-8℃，避光防潮保存。7. 有效期：6个月免责声明1. 豪运国际仅供研究使用，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. Human Collagen triple helix repeat-containing protein 1 (CTHRC1) ELISA Kit instruction Intended useThis CTHRC1 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 CTHRC1 in the sample, this CTHRC1 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 CTHRC1concentration. The concentration of CTHRC1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge 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.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.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 bags11Note: Standard (S0 → S5) concentration was followed by:0,25,50,100,200,400 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 step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis 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 ng/ml6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
HA-Tag Monoclonal Antibody(1B10)Catalog NoDM-Ab-04725IsotypeIgGReactivitySpecies independentApplicationsWB;ELISA;IP;IFGene NameHA TagProtein NameHA-TagImmunogenSynthetic Peptide of HA-TagSpecificityThe antibody detects C-terminal, internal, and N-terminal HA-tag fusion proteins.FormulationPBS, pH 7.4, containing 0.5%BSA, 0.02% sodium azide as Preservative and 50% Glycerol.SourceMonoclonal, MousePurificationThe antibody was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen.DilutionWB: 1:5000 IP: 1:200 IF: 1:1000;ELISA 1:5000-20000Concentration1 mg/mlPurity≥90%Storage Stability-20°C/1 yearSynonymsObserved BandCell PathwayTissue SpecificityFunction
CARD15/NOD2 rabbit pAb货号DM-Ab-18010同位型IgG应用WB种属Human靶点NOD2基因名称NOD2 CARD15 IBD1蛋白名称Nucleotide-binding oligomerization domain-containing protein 2 (Caspase免疫原Synthesized peptide derived from human CARD15/NOD2特异性This antibody detects endogenous levels of CARD15/NOD2 at Human组成Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.来源Polyclonal, Rabbit,IgG稀释WB 1:500-2000纯化工艺The antibody was affinity-purified from rabbit antiserum byaffinity-chromatography using epitope-specific immunogen.分子量114kD使用建议 This product can be used in immunological reaction related experiments. Formore information, please consult technical personnel.
IRF-5 Polyclonal AntibodyCatalog NoDM-Ab-01180IsotypeIgGReactivityHuman;MouseApplicationsWB;ELISAGene NameIRF5Protein NameInterferon regulatory factor 5 (IRF-5)ImmunogenSynthesized peptide derived from human protein . at AA range: 380-460SpecificityIRF5 Polyclonal Antibody detects endogenous levels of protein.FormulationLiquid in PBS containing 50% glycerol, and 0.02% sodium azide.SourcePolyclonal, Rabbit,IgGPurificationThe antibody was affinity-purified from rabbit antiserum byaffinity-chromatography using epitope-specific immunogen.DilutionWB 1:500-2000 ELISA 1:5000-20000Concentration1 mg/mlPurity≥90%Storage Stability-20°C/1 yearSynonymsObserved Band54kD
钙检测豪运国际(邻甲酚酞络合酮比色法)产品简介： 钙(Calcium)是一种金属元素， 常温下呈银白色晶体,动物的骨骼、蛤壳、蛋壳都含有碳 酸钙，检测生命体钙含量，主要通过检测钙离子浓度实现的，用分光度法检测钙较为常见， 该法需要合适的金属指示剂或选择性结合钙离子后引起变色的染料化合物，目前较为常用的 染料有邻-甲酚酞络合酮(OCPC)、偶氮砷Ⅲ、 甲基麝香草酚蓝(MTB)等。 钙检测豪运国际(邻甲酚酞络合酮比色法)是利用溶液中钙离子在碱性条件下能与OCPC 结合， 生成紫红色的络合物， 加入镁离子螯合剂， 去除镁离子背景干扰， 通过分光光度计 检测 575nm 处吸光度， 根据公式计算出总钙含量； 自备材料：离心管或试管、比色杯、分光光度计生化豪运国际组织样本前处理标准化流程一、核心原则快速处理：组织离体后立即操作或液氮速冻，防止蛋白酶、核酸酶降解目标物（蛋白、酶、代谢物等）。低温操作：全程保持 4℃或冰浴，降低生物分子活性损失。充分匀浆：破坏组织细胞膜结构，确保目标物充分释放。避免污染：耗材灭菌处理，核酸类检测需无酶环境，蛋白类检测避免蛋白酶污染。二、通用前处理步骤步骤操作要点注意事项组织取样与称重1. 取新鲜组织，用预冷的 PBS / 生理盐水冲洗表面血迹、杂质；2. 滤纸吸干水分，精确称取 50-200mg（根据豪运国际要求调整）1. 取样工具（剪刀、镊子）需预冷或灭菌；2. 避免反复冻融组织，建议分装保存组织匀浆制备1. 按 质量体积比 1:9 加入预冷的裂解液 / 提取液（如 100mg 组织 + 900μL 提取液）；2. 选择匀浆方式： - 机械匀浆：组织匀浆机 / 研磨器冰浴研磨至无明显颗粒； - 超声破碎：适用于坚硬组织（如肌肉、肝脏），功率适中避免产热； - 液氮研磨：适用于核酸、活性蛋白提取，研磨成粉末后加入提取液1. 匀浆过程全程冰浴，防止温度升高降解目标物；2. 超声时间不宜过长，避免蛋白变性离心分离1. 将匀浆液转移至离心管，4℃下 3000-12000rpm 离心 10-20min（转速和时间根据豪运国际及目标物调整）；2. 小心吸取上清液，避免触及沉淀（细胞碎片、细胞核1. 离心机提前预冷至 4℃；2. 上清液即为待测样本，若有浑浊可再次离心样本稀释与保存1. 根据豪运国际检测范围，用提取液 / 稀释液调整样本浓度；2. 即时检测：样本置于冰浴；3. 长期保存：分装后 - 20℃或 - 80℃冻存，避免反复冻融1. 稀释倍数需记录，用于＊终结果计算；2. 含酶样本建议添加酶抑制剂（如 PMSF）三、不同检测目标的针对性优化1.酶活性检测提取液需含对应酶保护剂（如激酶加 DTT，磷酸酶加磷酸酶抑制剂）；匀浆和离心步骤尽量缩短，减少酶活性损失；避免使用强变性剂（如 SDS）。2.蛋白质定量（BCA / 考马斯亮蓝法）若组织含高浓度脂肪 / 色素，需增加脱脂步骤（如用丙酮沉淀蛋白）；裂解液避免含强还原剂（如 β- 巯基乙醇），防止干扰显色反应。3.代谢物检测（糖、脂质、氨基酸）提取液选择适配豪运国际的专用缓冲液（如检测血糖用 Tris-HCl 缓冲液）；离心转速可提高至 12000rpm，确保彻底去除沉淀。4.核酸提取（DNA/RNA）液氮研磨后立即加入含 RNase 抑制剂的裂解液（RNA 提取）；避免使用金属器具，防止核酸降解；离心后上清液需进一步纯化（如酚氯仿抽提、柱纯化）。四、常见问题及解决方案常见问题原因解决方案上清液浑浊，杂质多匀浆不充分、离心转速过低延长匀浆时间，提高离心转速至 10000rpm 以上目标物含量过低组织取样量不足、裂解液比例不当增加组织取样量，调整质量体积比至 1:5~1:9酶活性检测结果偏低操作温度过高、匀浆时间过长全程冰浴操作，缩短匀浆和离心时间，添加酶抑制剂样本反复冻融后结果不稳定生物分子降解样本分装小体积冻存，避免反复冻融五、生化豪运国际使用全程注意事项实验前准备注意事项试剂管理严格按说明书温度解冻 / 平衡试剂：冷冻试剂（酶标物、标准品）需提前分装，避免反复冻融；冷藏试剂室温平衡 15-30 min，减少温度差对反应体系的影响。 检查试剂状态：若出现浑浊、沉淀、变色、异味，或超出有效期 / 开封后使用期限，立即废弃。 试剂混匀方式：解冻后轻柔颠倒混匀，禁止剧烈振荡，防止酶活性失活或产生气泡干扰检测。 样本预处理样本需澄清无杂质，浑浊样本 4℃ 3000-5000 rpm 离心 10-15 min 取上清，或 0.22 μm 滤膜过滤；脂质污染样本加 1% Triton X-100 处理后离心。 严格按说明书稀释样本，避免浓度超出检测线性范围；稀释用缓冲液需与豪运国际配套，禁止用水替代。 仪器与耗材准备校准酶标仪 / 分光光度计波长，用空白孔调零，确保仪器处于正常工作状态。 选用一次性无菌比色杯 / 酶标板，重复使用的玻璃器皿需彻底清洗并灭菌，避免残留洗涤剂或污染物干扰反应。 实验操作注意事项加样规范标记孔位（空白孔、标准品孔、样本孔、质控孔），避免混淆；建议按空白→标准品→质控品→样本的顺序加样。 使用校准过的移液器，控制加样体积＊＊度；不同孔位更换吸头，防止交叉污染。 加样时避免产生气泡，若有气泡可用移液器尖端轻轻刺破，或静置片刻待气泡消散。 孵育反应控制严格遵循说明书的孵育温度（如室温、37℃）和时间，温度误差控制在 ±1℃内，孵育时间不得随意延长或缩短。 避光反应需用锡箔纸包裹反应容器，防止光照导致底物分解；水浴孵育时确保反应容器完全浸入水中，避免边缘效应。 终止反应操作需终止反应的豪运国际，到达孵育时间后立即加入终止液，加液顺序和体积与说明书一致；加液后轻柔混匀，避免剧烈振荡。 实验后处理注意事项结果计算与验证绘制标准曲线时，确保 R²≥0.99；样本浓度需乘以稀释倍数，若超出标准曲线范围需重新稀释检测。 对比质控品检测值与说明书给定范围，若超出范围需排查原因（试剂、操作、仪器）并重新实验。 试剂与耗材处理未用完的试剂按组分特性分类保存（冷藏 / 冷冻 / 室温），密封瓶口并标注开封日期；冷冻试剂分装后避免再次冻融。 实验废液按生物安全要求处理，一次性耗材（吸头、酶标板）需高压灭菌后丢弃，防止污染环境。 数据记录与追溯完整记录实验信息：试剂批号、有效期、孵育条件、仪器参数、样本信息等，便于结果追溯和问题排查。相关产品：DM-FAM1020肝脂酶（HL）测试盒微量法DM-FAM1021肝脂酶（HL）测试盒可见分光光度法DM-FAM1022甘油三酯（TG）含量测试盒微量法DM-FAM1023甘油三酯（TG）含量测试盒可见分光光度法DM-FAM1024总胆固醇（TC）含量测试盒微量法DM-FAM1025总胆固醇（TC）含量测试盒可见分光光度法DM-FAM1026游离胆固醇（FC）含量测试盒微量法DM-FAM1027游离胆固醇（FC）含量测试盒可见分光光度法
鸡（Chicken）磷脂酰胆碱（PC）ELISA检测豪运国际本豪运国际只能用于科学研究，不得用于医学诊断检测原理豪运国际采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被磷脂酰胆碱（PC）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＊终的黄色。颜色的深浅和样品中的磷脂酰胆碱（PC）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分离。2. 血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。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-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1. 豪运国际保存在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，＊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。豪运国际组成名称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、30、60、120、240、480 pg/mL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60min。5. 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15min。7. 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 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。 豪运国际性能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：＊低检测浓度小于1.0 pg/mL。3. 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1. 豪运国际仅供研究使用，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。
鸡（Chicken）游离胆固醇（FC）ELISA检测豪运国际本豪运国际只能用于科学研究，不得用于医学诊断检测原理豪运国际采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被游离胆固醇（FC）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＊终的黄色。颜色的深浅和样品中的游离胆固醇（FC）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分离。2. 血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。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-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1. 豪运国际保存在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，＊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。豪运国际组成名称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、0.5、1、2、4、8 mmol/L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60min。5. 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15min。7. 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 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。 豪运国际性能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：＊低检测浓度小于0.1 mmol/L。3. 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1. 豪运国际仅供研究使用，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。
鸡（Chicken）黄体生成素（LH）ELISA检测豪运国际本豪运国际只能用于科学研究，不得用于医学诊断检测原理豪运国际采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被黄体生成素（LH）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＊终的黄色。颜色的深浅和样品中的黄体生成素（LH）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分离。2. 血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。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-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1. 豪运国际保存在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，＊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。豪运国际组成名称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、10、20、40、80、160 ng/ml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60min。5. 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15min。7. 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 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。 豪运国际性能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：＊低检测浓度小于1.0 ng/ml。3. 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1. 豪运国际仅供研究使用，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. Chicken Luteinizing Hormone (LH) ELISA Kit instruction Intended useThis LH 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 LH in the sample, this LH 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 LH concentration. The concentration of LH in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge 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.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.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 bags11Note: Standard (S0 → S5) concentration was followed by:0,10,20,40,80,160 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 step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis 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 ng/ml.6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
鸡（Chicken）双氢睾酮（DHT）ELISA检测豪运国际本豪运国际只能用于科学研究，不得用于医学诊断检测原理豪运国际采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被双氢睾酮（DHT）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＊终的黄色。颜色的深浅和样品中的双氢睾酮（DHT）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分离。2. 血浆：EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。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-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1. 豪运国际保存在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，＊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。豪运国际组成名称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、0.5、1、2、4、8 nmol/L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60min。5. 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15min。7. 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 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。 豪运国际性能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：＊低检测浓度小于0.1 nmol/L。3. 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1. 豪运国际仅供研究使用，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. Chicken Dihydro testosterone (DHT) ELISA Kit instruction Intended useThis DHT 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 DHT in the sample, this DHT 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 DHT concentration. The concentration of DHT in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge 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.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.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 bags11Note: Standard (S0 → S5) concentration was followed by:0,0.5,1,2,4,8 nmol/LReagent 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 step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis 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 0.1 nmol/L6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
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