人尿微量白蛋白(m-Alb)检测试剂盒(透射比浊法)
尿微量白蛋白(MAU)测定标准操作程序SOP文件
其它适合的质控品
贮存条件：置2-8℃冰箱至有效期。
准备：直接使用。
质控间隔时间及限制：应视不同地区及各自实验室情况而定。质控结果应在限定的范围之内，如果超出范围，实验室应根据情况采取措施。
ABCD医院
生化实验室
文件编号：
ABCD-SOP-04-13
10.5 MAU的浓度低于3000mg/l时不会有HOOK效应。
ABCD医院
生化实验室
文件编号：
ABCD-SOP-04-13
尿微量白蛋白（MAU）测定
版序：ABCD
页码：第3页，共3页
11临床意义
白蛋白是非糖类蛋白，分子量为66，000道而顿。其在肝实质细胞中合成，每天大约为14g/天。白蛋白是血浆、脑脊液、尿液中最重要的蛋白成份（约大于50％）。尿液中少量的但是非正常的白蛋白排泌称为尿微量白蛋白。
<37mg albumin /g crea
7.2 24-hour尿液：<20mg/l
<30mg/24h
8性能指标
本法线性范围为3-120mg/dl，不准确度允许范围 ±10%，不精密度血液：CV=3.3%，尿液：CV=4.3%灵敏度为1.5mg/dl。
9注意事项
9.1血清标本出现溶血、脂血、黄疸或抗坏血酸的干扰情况参见抗干扰能力。
微量白蛋白尿可以由小球、小管及肾后性等原因引起。白蛋白是不同的类型的蛋白尿的标记蛋白。
对于选择性的肾小球性蛋白尿，排泌量约为100－3000mg/g crea。对于非选择性的肾小球性蛋白尿，其特点为高分子量蛋白的分泌增加。肾前性蛋白尿通过总蛋白与白蛋白之间的不同来确认（白蛋白少于30％）。白蛋白及微量蛋白同事增加可见于球管性蛋白尿（肾小管的重吸收少于肾小球的滤过率）。肾小球、肾小管以及间质性肾炎、糖尿病或其它原因引起的肾衰。血浆中的蛋白质具有两种功能：维持渗透压以及参与体内物质的转运。对于低水溶性物质来说，白蛋白是最重要的转运蛋白。白蛋白水平的降低可以由高血压、肝细胞合成减少、血管内分泌异常、血管内外分布异常，白蛋白丢失及降解加快等原因引起。
尿微白检验方法学
尿微白检验方法学
尿微量白蛋白的检测方法主要有：ELISA方法、放射免疫法、化学发光法、免疫透射比浊法以及免疫荧光法。
ELISA方法灵敏性高，但操作步骤多，等待检测结果时间比较长，需要有专门的实验室、专业人员完成，一般用于大批量标本的检测。
放射免疫法灵敏度高、操作简便，测量和数据处理易于实现自动化。
但因其为相对定量及存在放射线辐射、污染等问题，所以应用较少。
化学发光法因其特异性、敏感性高、简便快速、无需外来光源、背景噪音低而应用广泛，但其选择性差、发射强度依赖于各种环境因素，需严格控制外界各种因素。
免疫透射比浊法试剂较稳定，自动化操作减少了人为误差，但其检测需核实的抗原抗体比例，需预先定值，否则会造成误差。
免疫荧光法特异性强、敏感性高、操作简便、检测时间短，即可批量检测也可单份检测，是临床上常用的快速定量检测方法。
白蛋白(ALB)测定试剂(盒)(溴甲酚绿法)产品技术要求新产业
白蛋白（ALB）测定试剂（盒）（溴甲酚绿法）
2.性能指标
2.1外观
试剂应为清澈透明的液体，无沉淀、悬浮物和絮状物。
2.2净含量
试剂装量的装量应按表 2，液体装量的最大允许负偏差应为 5%。
表 2 净含量
2.3试剂空白吸光度
试剂（盒）以纯化水为空白在 37 ℃±1℃ 、630 nm 波长、1 cm 光径条件下，试剂空白吸光度应≤0.40。
2.4分析灵敏度
试剂（盒）测试 43.7 g/L 的被测物时，吸光度变化（ΔA）应在 1.00～2.66 的范围内。
2.5线性范围
试剂（盒）在(0～60) g/L范围内的分析性能应符合如下要求：
a)线性相关系数r≥0.990；
b)(0～10) g/L 范围内，线性绝对偏差应在±1 g/L以内；(10～60) g/L 范围内，线性
相对偏差应在±10%以内。
2.6测量精密度
2.6.1重复性
用校准品重复测试所得结果的变异系数CV≤4%。
2.6.2批间差
试剂（盒）批间相对偏差R≤6%。
2.7准确度
校准品的相对偏差 B 在±10%以内。
2.8分析特异性
血红蛋白浓度在 400 mg/dL 内、甘油三酯浓度在 500 mg/dL 内、抗坏血酸浓度在 30 mg/dL 内、胆红素浓度在 40 mg/dL 内，对试剂检测结果的偏差影响应在±10%以内。
尿微量白蛋白定量检测操作程序
尿微量白蛋白定量检测操作程序1.目的规范尿微量白蛋白定量检测试验，确保检测结果准确性和重复性。
2．范围本操作规程适用于生化室工作人员、实习人员、进修人员的操作前培训。
3.术语4．测定原理4．1测定方法：免疫比浊法4．2 测定原理：样品中的微量自蛋白与特异性的抗人白蛋白抗体在液相中相遇，立即形成不溶性抗原-抗体复合物，并产生一定的浊度。
j虫度高低反映样品中MAlb的含量，通过与同样处理的校准品比较，即可计算出样品中MAlb的含量。
5．标本5．1标本种类：随机尿及24h尿，常温下1天，样品中MAlb在2~8°C保存可稳定l个月，-20·C保存可稳定6个月。
5．2标本采集：见生化标本采集程序。
6．试剂6．1试剂来源：新成生物有限公司。
6．2组成规格：R1 90ml*2 R2 45ml*16．3试剂准备本试剂为液体双试剂，可直接使用。
6．4试剂稳定性试剂2~8·C密闭避光贮存可稳定12个月，开瓶上机2~ 8°C避光贮存可稳定90天。
7. 检测程序7. 1试剂的准备：加入生化分析仪试剂盘中。
7. 2检测步骤：7. 2. 1基本参数方法:终点法主波长:340nm 样品/试剂:6/125ul 反应温度37·C 反应时间10min 副波长:700nm7. 2. 2校准：使用试剂盒配套的校准品；当试剂更换批号、质控出现漂移、仪器保养、重要零部件更换后应重新校准。
8.性能指标8.1、分析灵敏度：0mg/L。
8.2、测定范围：0~400mg/L。
8.3、精密度：批内cv.=5.0%、批间相对极差=10.0%。
8.4、准确度：相对偏差=10%。
8.5、空白眼光度：试剂空白在主波长340nm、副波长700nm处，10mm光径下，吸光度值<0.500。
9. 参考范围10. 临床意义10. 1 正常情况下，氨在肝脏转变成尿素。
严重肝脏疾病是，氨不能从循环中清除，引起血氨增高。
尿微量总蛋白 测定原理
尿微量总蛋白测定原理尿微量总蛋白（urine microalbumin）是指尿液中微量存在的白蛋白，通常用于评估肾脏疾病的早期诊断和监测疾病进展情况。
测定尿微量总蛋白的原理主要是利用免疫学方法，通过特定的试剂与蛋白质发生免疫相应，从而定量测定尿液中微量蛋白的浓度。
本文将从测定尿微量总蛋白的原理、常用的测定方法和临床应用等方面进行详细介绍。
一、测定原理尿微量总蛋白的测定原理主要是利用免疫学方法，即抗原与抗体的免疫相应原理。
通常采用免疫层析、免疫电泳、免疫荧光检测、免疫比浊法等方法进行测定。
1.免疫层析法免疫层析法是通过将尿样添加特定的抗体或抗原，使之与待测蛋白进行特异性结合，然后通过一系列步骤来分离和测定目标蛋白。
其原理是将试样通过一定具有特异性的固相支持介质，使之与固定的抗体或抗原结合，并随着尿样中存在的微量总蛋白一起被分离出来。
2.免疫电泳法免疫电泳法是将待测尿样加入到具有孔隙性的凝胶中，然后加上特异性的抗体或抗原，使之与待检蛋白结合，再通过电泳方法分离并测定蛋白的浓度。
3.免疫荧光检测法免疫荧光检测法是利用荧光标记的抗体或抗原与待测样品中的蛋白质特异结合，然后通过使用激光或紫外光源来激发荧光染料的发光信号，最后测定光信号的强度来计算出蛋白质的浓度。
4.免疫比浊法免疫比浊法是将待测尿样与特定的抗体或抗原结合，形成抗原-抗体复合物，然后通过测定溶液的浑浊度来计算蛋白质的浓度。
通过上述免疫学方法，可以快速、准确地测定尿液中微量总蛋白的浓度，具有较高的敏感性和特异性，能够有效的评估肾脏损伤的程度，为临床诊断和治疗提供重要依据。
二、常用的测定方法尿微量总蛋白的测定方法主要包括半定量法和定量法两种，其中定量法的测定结果更为准确。
1.半定量法半定量法通常采用尿液尿蛋白/肌酐比值进行测定，这是一种简便快速的半定量测定方法。
可以通过免疫比浊法或免疫电泳法等进行半定量分析，但由于其测定结果受尿液稀释度的影响较大，准确性较差。
尿微量白蛋白测定试剂盒(免疫比浊法)产品技术要求zhongshengbeikong
尿微量白蛋白测定试剂盒（免疫比浊法）适用范围：本试剂盒与ABBOTT ARCHITECT c4000/c8000/c16000全自动生化分析仪配套使用，用于体外定量测定人尿液中白蛋白的浓度。
1.1规格液体双剂型试剂1（R1)：60mL×2,试剂2(R2)： 6mL×2,校准品（4个浓度）：1mL×4/套；试剂1（R1)：65mL×2,试剂2(R2)： 10mL×2,校准品（4个浓度）：1mL×4/套。
1.2规格划分说明根据净含量划分规格。
1.3主要组成成分试剂盒由试剂1（R1）液体、试剂2（R2）液体及校准品液体组成。
1.3.1 试剂1（R1）液体主要组分：三羟甲基氨基甲烷（pH 7.5） 200mmol/L1.3.2 试剂2（R2）液体主要组分：羊抗人白蛋白抗体浓度根据效价而定1.3.3 校准品液体主要组分：PBS缓冲液基质（4个浓度）白蛋白①0 mg/L、②30mg/L～50mg/L、③70mg/L～100mg/L、④ 160mg/L～185mg/L。
（每批定值）2.1 外观试剂盒中各组件的外观应满足：a) 试剂1(R1)应为无色透明溶液，无杂质、无絮状物，外包装完整无破损；b) 试剂2(R2)应为浅黄色粘稠溶液，无杂质、无絮状物，外包装完整无破损；c) 校准品应为无色液体，无杂质、无絮状物，外包装完整无破损。
2.2 净含量液体试剂净含量应不少于标示值。
2.3 试剂空白吸光度在波长340nm处（光径1cm），试剂空白吸光度值（A）应≤0.2。
2.4 准确度用中生试剂和已上市同类试剂分别测定40个在测定范围内不同浓度的样本，在[20，180]mg/L检测范围内，比对两组数据的相关系数（r）及测值的偏差，要求r≥0.975，相对偏差应不超过±15％。
2.5 分析灵敏度对应于浓度为50mg/L的MALB所引起的吸光度变化差值（△A）的绝对值应在0.15～0.60的范围内。
尿微量白蛋白检测的标准操作程序
尿微量白蛋白检测的标准操作程序【目的】掌握NEPhstar特定蛋白分析仪检测尿微量白蛋白正确方法，保证检查项目的准确性。
【该SOP变动程序】本标准操作程序的改动，可由任一使用本SOP的工作人员提出，请专业组长及科主任签字后生效。
【检测原理】用散射比浊法，可溶性抗原与特异性的抗体反应形成不溶性复合物，当光线通过反应悬液时发生散射并由特种蛋白分析仪检测到。
散射光值的多少与测试样本中的蛋白浓度成一定比列。
通过刷卡将磁条中的标准曲线存储入仪器中，仪器会自动计算样品中的特定蛋白浓度。
【试剂组成】以上试剂由深圳国赛生物科技有限公司生产【所需其他材料】1．NEPHSTAR特定蛋白分析仪2．NEPHSTAR检测附件3．电子移液器YB2014．普通移液器【操作步骤】1、打开NEPHSTAR特定蛋白分析仪电源，仪器显示23、输入代码61，刷磁卡按刷磁卡，按ENTER直接进入下一步，默认仪器的序号和稀释度按此时4、将装有搅拌子和20ul尿液样品的测量杯放入测量室。
5移液器YB201将400ul缓冲液和40ul抗血清加入测量杯。
6、NEPHSTAR特定蛋白分析仪自动感应试剂的加入，搅拌子开始搅拌，测试开始。
测试完毕后仪器自动显示打印结果。
【参考区间】mALB<25mg/L【临床意义】1、正常情况下白蛋白在血液中检测出，并由肾脏过滤，当肾脏功能正常时白蛋白不会出现在尿液中，当肾小球基底膜受到损伤至通透性改变时，会有小部分白蛋白漏入尿液，形成微量白蛋白尿，高血压、糖尿病及系统性红斑狼疮等常伴有肾脏病变的缓慢进行性恶化，尿液中可较早发现异常。
2、尿液中白蛋白排泄量变动较大，随机尿标本一次尿微量白蛋白升高可能并无意义，如连续2-3次升高均超过参考区间方有诊断价值。
【注意事项】1、不要将批号的试剂要刷磁卡，磁卡里含有重要的标准曲线信息。
不同批号试剂不能混用。
2、试剂盒未开启前应存于2-8℃冰箱。
缓冲液和样品处理液在使用前放置室温平衡至18-26℃.抗血清和质控可稳定1个月。
cusabio 微量白蛋白尿(MAU ALB)检测试剂盒使用说明书
Human microalbunminuria(MAU/ALB) ELISA kit Catalog Number. CSB-E08970hFor the quantitative determination of human microalbunminuria(MAU/ALB) concentrations in serum, plasma, urine.This package insert must be read in its entirety before using this product.If You Have ProblemsTechnical Service Contact informationPhone: 86-27-87582341Fax: 86-27-87196150Email:****************Web: In order to obtain higher efficiency service, please ready to supply the lot numberof the kit to us (found on the outside of the box).1PRINCIPLE OF THE ASSAYThis assay employs the competitive inhibition enzyme immunoassay technique. Antibody specific for MAU has been pre-coated onto a microplate. Standards and samples are pipetted into the wells with biotin-conjugated MAU. A competitive inhibition reaction is launched between MAU (Standards or samples) and biotin-conjugated MAU with the pre-coated MAU antibody. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound reagent, a substrate solution is added to the wells and color develops in opposite to the amount of MAU bound in the initial step. The color development is stopped and the intensity of the color is measured.DETECTION RANGE0.078 µg/ml-5 µg/ml.SENSITIVITYThe minimum detectable dose of human MAU is typically less than 0.019 µg/ml. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest human MAU concentration that could be differentiated from zero.SPECIFICITYThis assay has high sensitivity and excellent specificity for detection of human MAU. No significant cross-reactivity or interference between human MAU and analogues was observed.Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human MAU and all the analogues, therefore, cross reaction may still exist.2PRECISIONIntra-assay Precision (Precision within an assay): CV%<8%Three samples of known concentration were tested twenty times on one plate to assess.Inter-assay Precision (Precision between assays):CV%<10%Three samples of known concentration were tested in twenty assays to assess.LIMITATIONS OF THE PROCEDUREFOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.The kit should not be used beyond the expiration date on the kit label.Do not mix or substitute reagents with those from other lots or sources.If samples generate values higher than the highest standard, dilute the samples with Sample Diluent and repeat the assay.Any variation in Sample Diluent, operator, pipetting technique, washing technique, incubation time or temperature, and kit age can cause variation in binding.This assay is designed to eliminate interference by soluble receptors, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.3MATERIALS PROVIDEDReagents QuantityAssay plate (12 x 8 coated Microwells) 1(96 wells) Standard (Freeze dried) 2Biotin-conjugate (100 x concentrate) 1 x 60 µlHRP-avidin (100 x concentrate) 1 x 120 µlBiotin-conjugate Diluent 1 x 10 mlHRP-avidin Diluent 1 x 20 ml Sample Diluent 2 x 20 mlWash Buffer (25 x concentrate) 1 x 20 mlTMB Substrate 1 x 10 mlStop Solution 1 x 10 ml Adhesive Strip (For 96 wells) 4Instruction manual 1STORAGEUnopenedkitStore at 2 - 8°C. Do not use the kit beyond the expiration date.Opened kitCoated assayplateMay be stored for up to 1 month at 2 - 8°C.Try to keep it in a sealed aluminum foil bag,and avoid the damp.Standard May be stored for up to 1 month at 2 - 8° C.If don’t make recent use, better keep it storeat -20°C.HRP-avidinBiotin-conjugateBiotin-conjugateDiluentMay be stored for up to 1 month at 2 - 8°C. HRP-avidinDiluentSample DiluentWash BufferTMB SubstrateStop Solution*Provided this is within the expiration date of the kit.4OTHER SUPPLIES REQUIREDMicroplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.An incubator which can provide stable incubation conditions up to 37°C±0.5°C.Squirt bottle, manifold dispenser, or automated microplate washer.Absorbent paper for blotting the microtiter plate.100ml and 500ml graduated cylinders.Deionized or distilled water.Pipettes and pipette tips.Test tubes for dilution.PRECAUTIONSThe Stop Solution provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.5SAMPLE COLLECTION AND STORAGESerum Use a serum separator tube (SST) and allow samples to clot for30 minutes before centrifugation for 15 minutes at 1000 x g, 2 - 8°C.Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Centrifuge the sample again after thawing before the assay.Plasma Collect plasma using EDTA, or heparin as an anticoagulant.Centrifuge for 15 minutes at 1000 x g, 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Centrifuge the sample again after thawing before the assay.Urine Use a sterile container to collect urine samples. Remove any particulates by centrifugation for 15 minutes at 1000xg, 2 - 8°C and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Centrifuge again before assaying to remove any additional precipitates that may appear after storage.SAMPLE PREPARATIONRecommend to dilute the serum or plasma samples 50000-fold before test.The suggested 50000-fold dilution can be achieved by adding 2µl sample to 398µl of normal saline. Complete the 50000-fold dilution by adding 2µl of this solution to 498µl of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments.Recommend to dilute the urine samples with Sample Diluent(1:40) before test. The suggested 40-fold dilution can be achieved by adding 6µl sample to 234µl of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments6Note:1. CUSABIO is only responsible for the kit itself, but not for the samplesconsumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.2. Samples to be used within 5 days may be stored at 2-8°C, otherwisesamples must be stored at -20°C (≤1month) or -80°C (≤2month) to avoid loss of bioactivity and contamination.3. Grossly hemolyzed samples are not suitable for use in this assay.4. If the samples are not indicated in the manual, a preliminary experiment todetermine the validity of the kit is necessary.5. Please predict the concentration before assaying. If values for these arenot within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.6. Tissue or cell extraction samples prepared by chemical lysis buffer maycause unexpected ELISA results due to the impacts of certain chemicals.7. Owing to the possibility of mismatching between antigen from otherresource and antibody used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.8. Influenced by the factors including cell viability, cell number and alsosampling time, samples from cell culture supernatant may not be detected by the kit.9. Fresh samples without long time storage are recommended for the test.Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.7REAGENT PREPARATIONNote:Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit. Bring all reagents to room temperature (18-25°C) before use for 30min.Prepare fresh standard for each assay. Use within 4 hours and discard after use.Making serial dilution in the wells directly is not permitted.Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved.To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µl for once pipetting.Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.1. Biotin-conjugate (1x) - Centrifuge the vial before opening.Biotin-conjugate requires a 100-fold dilution. A suggested 100-fold dilution is 10 µl of Biotin-conjugate + 990 µl of Biotin-conjugate Diluent.2. HRP-avidin (1x) - Centrifuge the vial before opening.HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 µl of HRP-avidin + 990 µl of HRP-avidin Diluent.3. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up toroom temperature and mix gently until the crystals have completely dissolved. Dilute 20 ml of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 ml of Wash Buffer (1 x).894.StandardCentrifuge the standard vial at 6000-10000rpm for 30s.Reconstitute the Standard with 1.0 ml of Sample Diluent . Do not substitute other diluents. This reconstitution produces a stock solution of 5 µg/ml. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.Pipette 150 µl of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (5 µg/ml). Sample Diluent serves as the zero standard (0 µg/ml).Tube S7 S6 S5S4 S3 S2 S1 S0 µg/ml52.51.250.6250.3120.1560.078ASSAY PROCEDUREBring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay.It is recommended that all samples and standards be assayed in duplicate.1. Prepare all reagents, working standards, and samples as directed in theprevious sections.2. Refer to the Assay Layout Sheet to determine the number of wells to beused and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.3. Set a Blank well without any solution.4. Add 50µl of standard and sample per well.5. Add 50µl of Biotin-conjugate(1x) to each well(not to Blank well). Coverwith a new adhesive strip. Incubate for 60 minutes at 37°C.(Biotin-conjugate(1x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)6. Aspirate each well and wash, repeating the process two times for a total ofthree washes. Wash by filling each well with Wash Buffer (200µl) using a squirt bottle, multi-channel pipette, manifold dispenser, or autowasher, and let it stand for 2 minutes, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.7. Add 100µl of HRP-avidin(1x) to each well(not to Blank well). Cover themicrotiter plate with a new adhesive strip. Incubate for 60 minutes at 37°C.8. Repeat the aspiration/wash process for five times as in step 6.9. Add 90µl of TMB Substrate to each well. Incubate for 20 minutes at 37°C.Protect from light.10. Add 50µl of Stop Solution to each well, gently tap the plate to ensurethorough mixing.1011. Determine the optical density of each well within 5 minutes, using amicroplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.*Samples may require dilution. Please refer to Sample Preparation section. Note:1. The final experimental results will be closely related to validity of theproducts, operation skills of the end users and the experimental environments.2. Samples or reagents addition: Please use the freshly prepared Standard.Please carefully add samples to wells and mix gently to avoid foaming. Do not touch the well wall as possible. For each step in the procedure, total dispensing time for addition of reagents or samples to the assay plate should not exceed 10 minutes. This will ensure equal elapsed time for each pipetting step, without interruption. Duplication of all standards and specimens, although not required, is recommended. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions.Also, use separate reservoirs for each reagent.3. Incubation: To ensure accurate results, proper adhesion of plate sealersduring incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. Incubation time and temperature must be observed.4. Washing: The wash procedure is critical. Complete removal of liquid ateach step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting and remove any drop of water and fingerprint on the bottom of the plate. Insufficient washing will result in poor precision and falsely elevated absorbance reading. When using an automated plate washer, adding a 30 second soak period following the addition of wash buffer, and/or rotating the plate 180 degrees between wash steps may improve assay precision.115. Controlling of reaction time: Observe the change of color after adding TMBSubstrate (e.g. observation once every 10 minutes), TMB Substrate should change from colorless or light blue to gradations of blue. If the color is too deep, add Stop Solution in advance to avoid excessively strong reaction which will result in inaccurate absorbance reading.6. TMB Substrate is easily contaminated. TMB Substrate should remaincolorless or light blue until added to the plate. Please protect it from light.7. Stop Solution should be added to the plate in the same order as the TMBSubstrate. The color developed in the wells will turn from blue to yellow upon addition of the Stop Solution. Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the TMB Substrate.1213ASSAY PROCEDURE SUMMARY*Samples may require dilution. Please refer to Sample Preparation section.CALCULATION OF RESULTSUsing the professional soft "Curve Expert" to make a standard curve is recommended, which can be downloaded from our web.Average the duplicate readings for each standard and sample and subtract the average optical density of Blank.Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the MAU concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.14人尿微量白蛋白(MAU/ALB)酶联免疫试剂盒使用说明书【产品编号】CSB-E08970h【预期应用】ELISA法定量测定人血清、血浆、尿液中MAU含量。
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人尿微量白蛋白(m-Alb)检测试剂盒(透射比浊法)
简介：
Viberti 等人发现尿中白蛋白排泄的增加率，可以提示胰岛素依赖性糖尿病肾病发作，但此时尿中总蛋白排泄正常，尿常规蛋白定性实验为阴性，而尿白蛋白排泄增加，称之为微量白蛋白(microablumin ，m-Alb)尿。
Leagene 人尿微量白蛋白(m-Alb)检测试剂盒(透射比浊法)其检测原理是尿液中的白蛋白与抗白蛋白特异抗体在液相中反应生成m-Alb 抗原抗体复合物，使反应液呈一定浊度。
本试剂盒专门用于人尿液中的微量白蛋白含量测定。
本试剂盒仅用于科研领域，不宜用于临床诊断或其他用途。
组成：
操作步骤(仅供参考)：
1、 准备试剂：临用前，各种试剂均应平衡至室温。
2、 绘制标准曲线：取白蛋白标准(198mg/L)，依次稀释后供标准曲线用，其浓度分别为
198mg/L 、99.0mg/L 、49.5mg/L 、19.8mg/L 、9.9mg/L 。
3、 m-Alb 测定操作：分光光度计手工或半自动生化分析仪检测，按下表依次加入试剂：
4、 充分混匀, 盖上塑料膜，37℃孵育。
5、 再次混匀，分光光度计测定340nm 波长处的吸光度。
以空白管调零，读取标准管和各
待测管的最终吸光度为A 2。
6、 绘制标准曲线：以上述5种浓度的标准液，分别制作5个标准管，同上操作，测定吸
编号 名称
TC068950T Storage
试剂(A): m-Alb buffer 50ml RT 试剂(B): 白蛋白标准(198mg/L) 0.2ml -20℃ 试剂(C): m-Alb 抗体工作液 0.5ml -20℃ 试剂(D): m-Alb 生理盐水 30ml
RT 使用说明书
1份
标准管 待测管 m-Alb buffer(ml) 1.0 1.0 白蛋白标准(μl) 100 待测样本(μl)
100
混匀，m-Alb 生理盐水调零，读取起始吸光度A 1，然后加入： m-Alb 抗体工作液(μl)
100
100
光度。
A标准=A2标准-A1标准
A标准和对应的白蛋白浓度在半对数纸上制图，绘制标准曲线。
计算：A样本=A2样本-A1样本以A样本查标准曲线，即求得尿中白蛋白浓度。
参考区间：
健康成年人尿液白蛋白：
24h尿：<30mg/ 24h。
定时尿：<20μg/min。
随意尿：<30μg/mg肌酐。
注意事项：
1、上述试剂避免反复冻融，以免失效或效率下降。
2、本法属于浊度反应，试剂中有可见混浊出现后，应弃用。
3、如果没有分光光度计，也可以使用酶标仪测定。
使用酶标仪测定蛋白浓度时，每个试
剂盒可以测定的样品数量可能会显著增加。