兔子端粒酶ELISA试剂盒说明书

srgn elisa试剂盒使用说明书SRGN Elisa试剂盒使用说明书一、引言SRGN Elisa试剂盒是一种用于检测人体内SRGN（serglycin）蛋白含量的工具。

SRGN是一种分布广泛的天然蛋白质，在多种细胞类型中表达，并参与多种生物学过程。

本文将介绍SRGN Elisa试剂盒的使用方法和注意事项。

二、试剂盒组成SRGN Elisa试剂盒包括以下组成部分：1. 微孔板：包含已涂层的SRGN抗体。

2. 标准品：一系列已知浓度的SRGN标准品。

3. 反应液：包括洗涤缓冲液、稀释液、显色液和停止液。

4. 底物：TMB底物。

三、试剂盒的使用步骤1. 取出所需微孔板的数量，将多余的微孔板密封保存。

2. 向每个孔中加入100μL稀释液，轻轻摇晃，将孔底液完全溶解。

3. 倒出孔内的液体，用洗涤缓冲液冲洗孔底3次，每次加入300μL，倒出液体。

4. 向每个孔中分别加入50μL标准品或待测样本，注意避免交叉污染。

5. 加入100μL SRGN抗体，轻轻摇晃，保证孔底液完全溶解。

6. 将微孔板密封，放置于37℃恒温箱中孵育1小时。

7. 倒出孔内的液体，用洗涤缓冲液冲洗孔底3次，每次加入300μL，倒出液体。

8. 向每个孔中加入100μL显色液，轻轻摇晃，保证显色液均匀分布。

9. 将微孔板密封，放置于室温下避光孵育15分钟。

10. 加入50μL停止液，轻轻摇晃。

11. 使用Elisa阅读器在450nm波长下测量各孔的吸光度值。

四、结果分析根据吸光度值绘制标准曲线，计算待测样本中SRGN的浓度。

根据吸光度值和标准曲线的关系，可以通过软件或手动计算得出结果。

五、注意事项1. 试剂盒的保存应在2-8℃下，避免阳光直射和冻结。

2. 打开试剂盒后，应尽快使用，避免试剂的长时间接触空气。

3. 使用试剂盒前，将所有试剂和标本恢复至室温。

4. 各步骤中的孔内液体倒出时，应使用吸头或洗涤器，避免污染。

5. 实验过程中，避免试剂的接触皮肤和眼睛。

Rat IL-1beta ELISA KitCatalog NO.:RK00009version:2.0This package insert must be read in its entirety beforeusing this productIntroductionThe kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of IL-1beta in rat serum,plasma,cell culture supernatants.Principle of the AssayThis assay employs the quantitative sandwich enzyme immunoassay technique.An antibody specific for rat IL-1beta has been pre-coated onto a microplate.Standards and samples are pipetted into the wells and any IL-1beta present is bound by the immobilized antibody.After washing away any unbound substances, and then a detection antibody specific for IL-1beta is added to the wells and binds to the combination of capture antibody IL-1beta in sample.Following a wash to remove any unbound combination,and enzyme conjugate is added to the wells. Following incubation and wash steps,a substrate solution is added to the wells and color develops in proportion to the amount of IL-1beta bound in the initial step.The color development is stopped and the absorbance is measured.Material Provided&Storage ConditionsUnopened kits can be stored at2-8°C for1year,and opened products must be used within1month.Part Size Cat.No.Storage ofopened/reconstituted materialAntibody Coated Plate 8×12RM00064Put the unused slats backin the aluminum foil bagwith the desiccant andreseal them.They can bestored at2-8°C for1month.Standard Lyophilized 2vials RM00061It is not recommended touse again afterredissolving.Concentrated BiotinConjugate Antibody(100×)1×120ul RM00062Store at2-8°c for1month*Streptavidin-HRP Concentrated(100×)1×120ul RM00063Store at 2-8°c for 1month *Standard/SampleDiluent (R1)1×20mL RM00023Store at 2-8°c for 1month *Biotin-Conjugate AntibodyDiluent (R2)1×12mL RM00024Streptavidin-HRP Diluent(R3)1×12mL RM00025WashBuffer(20x)1×30mL RM00026TMB Substrate1×12mL RM00027Stop Solution1×6mL RM00028Plate Sealers4Strips Specification 1Other Supplies Required1.Microplate reader capable of measuring absorbance at450nm,with the correction wavelength set at630nm or570 nm.2.Pipettes and pipette tips.3.Deionized or distilled water.4.Squirt bottle,manifold dispenser,or automated microplatewasher.5.Incubator.6.Test tubes for dilution of standards and samples.Precautions1.Any variation in diluent,operator,pipetting technique,washing technique,incubation time or temperature,and kitage can cause variation in binding.2.Variations in sample collection,processing,and storagemay cause sample value differences.3.Reagents may be harmful,if ingested,rinse it with anexcess amount of tap water.4.Stop Solution contains strong acid.Wear eye,hand,andface protection.5.Please perform simple centrifugation to collect the liquidbefore use.6.Do not mix or substitute reagents with those from other lotsor other sources.7.Adequate mixing is particularly important for good result.Use a mini-vortexer at the lowest frequency.8.Mix the sample and all components in the kits adequately,and use clean plastic container to prepare all diluents.9.Both the sample and standard should be assayed in duplicate,and reagents should be added in sequence in accordance with the requirement of the specification.10.Reuse of dissolved standard is not recommended.11.The kit should not be used beyond the expiration date onthe kit label.12.The kit should be away from light when it is stored orincubated.13.To reduce the likelihood of blood-borne transmission ofinfectious agents,handle all serum,plasma,and other biological fluids in accordance with NCCLS regulations.14.To avoid cross contamination,please use disposablepipette tips.15.Please prepare all the kit components according to theSpecification.If the kits will be used several times, please seal the rest strips and preserve with desiccants.Do use up within2months.16.This assay is designed to eliminate interference by otherfactors present in biological samples.17.Until all factors have been tested in this assay,thepossibility of interference cannot be excluded.18.The48T kit is also suitable for the specification.Sample Collection&StorageThe sample collection and storage conditions listed below are intended as general guidelines.Sample stability has not been evaluated.Samples containing the correlated IgG as in this kit may interfere with this assay.Cell Culture Supernatant:Remove particulates by centrifugation. Assay immediately or aliquot and store samples at≤-20°C. Avoid repeated freeze-thaw cycles.Serum:Use a serum separator tube(SST)and allow samples to clot for30minutes at room temperature before centrifugation for15 minutes at1000x g.Remove serum and assay immediately or aliquot and store samples at≤-20°C.Avoid repeated freeze-thaw cycles.Plasma:Collect plasma using EDTA or Heparin as an anticoagulant. Centrifuge for15minutes at1000×g within30minutes of collection.Assay immediately or aliquot and store samples at ≤-20℃.Avoid repeated freeze-thaw cycles.(Note:Citrateplasma has not been validated for use in this assay.)Note:It is suggested that all samples in one experiment be collected at the same time of the day.Avoid hemolytic and hyperlipidemia sample for serum and plasma.Reagent PreparationBring all reagents to room temperature before use.If crystals have formed in the concentrate,Bring the reagent to room temperature and mix gently until the crystals have completely dissolved.Standard-Reconstitute the Standard Lyophilized with 1.0mL Standard/Sample Diluent(R1).This reconstitution produces a stock solution of8000pg/mL.Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15minutes with gentle agitation prior to making dilutions. Use the8000pg/mL standard stock to produce a dilution series (below)with Standard/Sample Diluent(R1).Mix each tube thoroughly and change pipette tips between each transfer(recommended concentration for standard curve:4000,2000,1000,500,250,125,62.5,0pg/mL).Use diluted standards within 60minutes of preparation.Working Biotin Conjugate Antibody -Dilute 1:100of Concentrated Biotin Conjugate Antibody (100x)with Biotin-Conjugate Antibody Diluent (R2)before use,for example:Add 20μL of Concentrated Biotin Conjugate Antibody (100x)to 1980μL Biotin-Conjugate Antibody Diluent (R2)to prepare 2000μL Working Biotin Conjugate Antibody Buffer.Working Streptavidin-HRP -Dilute 1:100of Concentrated Streptavidin-HRP (100x)with Streptavidin-HRP Diluent (R3)before use,for example:Add 20μL of Concentrated Streptavidin-HRP (100x)to 1980μL Streptavidin-HRP Diluent (R3)Std 250μL 250μL 250μL 250μL 250μL 250μL 250μL 250μL 1000pg/mL R11000μL 8000pg/mL 250μL 4000pg/mL 250μL 250pg/mL 250μL 500pg/mL 250μL 125pg/mL 250μL 2000pg/mL 250μL 62.5pg/mL 250μL 0pg/mLto prepare2000μL Working Streptavidin-HRP Buffer.Wash Buffer-If crystals have formed in the concentrate,warm to room temperature and mix gently until the crystals have completely dissolved.Dilute1:20with double distilled or deionized water before use,for example:Add20mL of Wash Buffer Concentrate to380mL of deionized or distilled water to prepare 400mL of Wash Buffer.Assay ProcedureBring all reagents and samples to room temperature before use. It is recommended that all standards,controls,and samples be assayed in duplicate.1.Prepare all reagents,working standards,and samples asdirected in the previous sections.2.Remove excess microplate strips from the plate frame,return them to the foil pouch containing the desiccant pack, and reseal properly.3.Add wash buffer350μL/well,aspirate each well afterholding40seconds,repeating the process two times fora total of three washes.4.Add100μL Standard/sample Diluent(R1)in a blank well.5.Add100μL different concentration of standard or samplein other wells,Cover with the adhesive strip provided.Incubate for2hours at37℃.record the plate layout ofstandards and sample assay.6.Prepare the Concentrated Biotin Conjugate Antibody(100x)Working Solution15minutes early before use.7.Repeat the aspiration/wash as in step 3.8.Add100μL Working Biotin Conjugate Antibody in each well,cover with new adhesive Sealer provided.Incubate for1hour at37℃.9.Prepare the Streptavidin-HRP Concentrated(100x)WorkingSolution15minutes early before use.10.Repeat the aspiration/wash as in step 3.11.Add100μL Working Streptavidin-HRP in each well,coverwith new adhesive Sealer provided.Incubate for0.5hourat37℃.12.Repeat the aspiration/wash as in step 3.13.During the incubation,turn on the microplate reader towarm up for30minutes before measuring.14.Add100μL TMB Substrate to each well.Incubate for15-20minutes at37℃.Protect from light.15.Add50μL Stop Solution,determine the optical density ofeach well within5minutes,using a Microplate reader setto450nm.If wavelength correction is available,set to570nm or630nm.If wavelength correction is not available, subtract readings at570nm or630nm from the readingsat450nm.This subtraction will correct for opticalimperfections in the plate.Readings made directly at450nm without correction may cause higher value and lessaccurate result.Assay Procedure SummaryPrepare the standard and reagentsWash3times↓Add100ul of standards or test samples to each well Incubate for2hours at37℃,then wash3times↓Add100ul Working Biotin Conjugate AntibodyIncubate for1hour at37℃,then wash3times↓Add100ul Working Streptavidin-HRPIncubate for0.5hour at37℃,then wash3times↓Add100ul Substrate SolutionIncubate for15-20min at37℃under dark condition↓Add50ul Stop Solution↓Detect the optical density within5minutes under450nm.Correction Wavelength set at570nm or630nmCalculation of Results1.Average the duplicate readings for each standard,controland sample,and subtract the average zero standard optical density(O.D.).2.Create a standard curve by reducing the data using computersoftware 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 Y-axis against the concentration on the X-axis and draw a best fit curve through the points on a log/log graph.The data may be linearized by plotting the log of the IL-1 beta concentrations versus the log of the O.D.on a linear scale,and the best fit line can be determined by regression analysis.3.If samples have been diluted,the concentration read fromthe standard curve must be multiplied by the dilution factor.Typical DataThe standard curves are provided for demonstration only.A standard curve should be generated for each set of IL-1beta assayed.Detection Range62.5-4000pg/mLSensitivityThe minimum detectable dose(MDD)of IL-1beta typically less than13.22pg/mL.The MDD was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.SpecificityThis assay recognizes both recombinant and natural rat IL-1beta. The factors listed below were prepared at50ng/ml and assayed for cross-reactivity.No significant cross-reactivity was observed with the following:Recombinant rat:Recombinant human:IL-10IL-1IFN-γIL-2IL-1αIL-6IL-2IL-4IL-6IL-8TNF-αNote:Limited by current skills and knowledge,it is impossible for us to complete the cross-reactivity detection between IL-1beta and all the analogues,therefore,cross reaction may still exist.PrecisionIntra-plate Precision3samples with low,middle and high level IL-1beta were tested 20times on one plate,respectively.Intra-Assay:CV<10%Inter-plate Precision3samples with low,middle and high level IL-1beta were tested on3different plates,20replicates in each plate.Inter-Assay:CV<15%Intra-Assay Precision Inter-Assay Precision Sample123123 n202020202020 Mean(pg/mL)116522683535116022593542 Standard deviation47.879.4137.975.4162.6219 CV(%) 4.1 3.5 3.9 6.57.2 6.2RecoveryMatrices listed below were spiked with certain level of IL-1beta and the recovery rates were calculated by comparing the measured value to the expected amount of IL-1beta in samples.Sample Average Recovery（%）Range（%）Cell Culture Media(n=5)10495-113 Serum(n=5)9990-108LinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of IL-1beta and their serial dilutions.The results were demonstrated by the percentage of calculated concentration to the expected.Cell Culture Media(n=5)Serum(n=5) //Average of Expected（%）103951:2Range（%）98-10887-102Average of Expected（%）981021:4Range（%）84-11189-114 Average of Expected（%）961001:8Range（%）86-10692-108Average of Expected（%）981061:16Range（%）91-10596-115Trouble ShootingProblem Possible Cause SolutionHigh Background Insufficient washingSufficiently wash plates asrequired.Ensure appropriateduration and number of washes.Ensure appropriate volume of washbuffer in each well.Incorrect incubationprocedureCheck whether the duration andtemperature of incubation are set upas required.Cross-contamination ofsamples and reagentsBe careful of the operations thatcould cause cross-contamination.Use fresh reagents and repeat thetests.No signal or weak signal Incorrect use ofreagentsCheck the concentration anddilution ratio of reagents.Makesure to use reagents in properorder.Incorrect use ofmicroplate readerWarm the reader up before use.Makesure to set up appropriate mainwavelength and correctionwavelength.Insufficient colourreaction timeOptimum duration of colour reactionshould be limited to15-25minutes.Read too late after stopping the colour reaction Read the plate in5minutes after stopping the reaction.Matrix effect ofsamplesUse positive control.Too much signal Contamination of TMBsubstrateCheck if TMB substrate solutionturns e new TMB substratesolution.Plate sealers reusedUse a fresh new sealer in each stepof experiments.Protein concentrationin sample is too highDo pre-test and dilute samples inoptimum dilution ratio.Poor Duplicates Uneven addition ofsamplesCheck the pipette.Periodicallycalibrate the pipette.Impurities andprecipitates insamplesCentrifuge samples before use.Inadequate mixing ofreagentsMix all samples and reagents wellbefore loading.*For research purposes only.Not for therapeutic or diagnostic purposes.。

蛙皮素6-14(BB)ELISA试剂盒说明书本试剂仅供研究使用标本：体液蛙皮素6-14(BB)ELISA试剂盒说明书试验原理：BFGF试剂盒是固相夹心法酶联免疫吸附实验（ELISA）.已知BFGF浓度的标准品、未知浓度的样品加入微孔酶标板内进行检测。

先将BFGF和生物素标记的抗体同时温育。

人碱性成纤维细胞生长因子ELISA 检测试剂盒洗涤后，加入亲和素标记过的HRP。

再经过温育和洗涤，去除未结合的酶结合物，然后加入底物A、B，和酶结合物同时作用。

产生颜色。

颜色的深浅和样品中BFGF的浓度呈比例关系。

蛙皮素6-14(BB)ELISA试剂盒说明书自备材料1．蒸馏水。

2．加样器：5ul、10ul、50ul、100ul、200、500ul、1000ul。

3．振荡器及磁力搅拌器等。

安全性1．避免直接接触终止液和底物A、B。

一旦接触到这些液体，请尽快用水冲洗。

2．实验中不要吃喝、抽烟或使用化妆品。

3．不要用嘴吸取试剂盒里的任何成份。

蛙皮素6-14(BB)ELISA试剂盒说明书操作注意事项1．试剂应按标签储存，使用前恢复到室温。

稀稀过后的标准品应丢弃，不可保存。

2．实验中不用的板条应立即放回包装袋中，密封保存，以免变质。

3．不用的其它试剂应包装好或盖好。

不同批号的试剂不要混用。

保质前使用。

4．使用一次性的吸头以免交叉污染，吸取终止液和底物A、B液时，避免使用带金属部分的加样器。

5．使用干净的塑料容器配置洗涤液。

使用前充分混匀试剂盒里的各种成份及样品。

6．洗涤酶标板时应充分拍干，不要将吸水纸直接放入酶标反应孔中吸水。

7．底物A应挥发，避免长时间打开盖子。

底物B对光敏感，避免长时间暴露于光下。

避免用手接触，有毒。

实验完成后应立即读取OD值。

8．加入试剂的顺序应一致，以保证所有反应板孔温育的时间一样。

9．按照中标明的时间、加液的量及顺序进行温育操作。

蛙皮素6-14(BB)ELISA试剂盒说明书样品收集、处理及保存方法1、血清-----操作过程中避免任何细胞刺激。

兔子谷胱甘肽(GSH)酶联免疫分析试剂盒使用说明书本试剂盒仅供研究使用。

检测范围：96T12 pg/ml-450 pg/ml使用目的：本试剂盒用于测定兔子血清、血浆及相关液体样本中谷胱甘肽(GSH)含量。

实验原理本试剂盒应用双抗体夹心法测定标本中兔子谷胱甘肽(GSH)水平。

用纯化的兔子谷胱甘肽(GSH)抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入谷胱甘肽(GSH)，再与HRP标记的谷胱甘肽(GSH)抗体结合，形成抗体-抗原-酶标抗体复合物，经过彻底洗涤后加底物TMB显色。

TMB在HRP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谷胱甘肽(GSH)呈正相关。

用酶标仪在450nm波长下测定吸光度（OD值），通过标准曲线计算样品中兔子谷胱甘肽(GSH)浓度。

试剂盒组成标本要求1.标本采集后尽早进行提取，提取按相关文献进行，提取后应尽快进行实验。

若不能马上进行试验，可将标本放于-20℃保存，但应避免反复冻融2.不能检测含NaN3的样品，因NaN3抑制辣根过氧化物酶的（HRP）活性。

操作步骤1. 标准品的稀释：本试剂盒提供原倍标准品一支，用户可按照下列图表在小试管中进行稀释。

2. 加样：分别设空白孔（空白对照孔不加样品及酶标试剂，其余各步操作相同）、标准孔、待测样品孔。

在酶标包被板上标准品准确加样50μl，待测样品孔中先加样品稀释液40μl，然后再加待测样品10μl（样品最终稀释度为5倍）。

加样将样品加于酶标板孔底部，尽量不触及孔壁，轻轻晃动混匀。

3. 温育：用封板膜封板后置37℃温育30分钟。

4. 配液：将30倍浓缩洗涤液用蒸馏水30倍稀释后备用5. 洗涤：小心揭掉封板膜，弃去液体，甩干，每孔加满洗涤液，静置30秒后弃去，如此重复5次，拍干。

6. 加酶：每孔加入酶标试剂50μl，空白孔除外。

7. 温育：操作同3。

8. 洗涤：操作同5。

9. 显色：每孔先加入显色剂A50μl，再加入显色剂B50μl，轻轻震荡混匀，37℃避光显色10分钟.。