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Optimization of LC-MS peptide mapping
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2005-08-25 00:00 450阅读次数
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Optimization of LC-MS peptide mapping
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Optimization of LC-MS peptide mapping Optimization of LC-MS peptide mapping[详细]
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大豆多肽（peptide）酶联免疫分析试剂盒使用说明书 大豆多肽（peptide）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：96T2ng/L-90ng/L使用目的：本试剂盒用于测定发酵豆粕样本中大豆多肽（peptide）含量。实验原理本试剂盒应用双抗体夹心法测定标本中大豆多肽（peptide）水平。用纯化的大豆多肽（peptide）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大豆多肽（peptide），再与HRP标记的大豆多肽（peptide）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Z终的黄色。颜色的深浅和样品中的大豆多肽（peptide）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大豆多肽（peptide）浓度。大豆多肽（peptide）酶联免疫分析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NaN3YZ辣根过氧化物酶的（HRP）活性。大豆多肽（peptide）酶联免疫分析试剂盒操作步骤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Z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15分钟.10.终止：每孔加终止液50μl，终止反应（此时蓝色立转黄色）。11.测定：以空白空调零，450nm波长依序测量各孔的吸光度（OD值）。测定应在加终止液后15分钟以内进行。大豆多肽（peptide）酶联免疫分析试剂盒操作程序总结：计算以标准物的浓度为横坐标，OD值为纵坐标，在坐标纸上绘出标准曲线，根据样品的OD值由标准曲线查出相应的浓度；再乘以稀释倍数；或用标准物的浓度与OD值计算出标准曲线的直线回归方程式，将样品的OD值代入方程式，计算出样品浓度，再乘以稀释倍数，即为样品的实际浓度。注意事项1．试剂盒从冷藏环境中取出应在室温平衡15-30分钟后方可使用，酶标包被板开封后如未用完，板条应装入密封袋中保存。2．浓洗涤液可能会有结晶析出，稀释时可在水浴中加温助溶，洗涤时不影响结果。3．各步加样均应使用加样器，并经常校对其准确性，以避免试验误差。一次加样时间**控制在5分钟内，如标本数量多，推荐使用排枪加样。4．请每次测定的同时做标准曲线，**做复孔。如标本中待测物质含量过高（样本OD值大于标准品孔**孔的OD值），请先用样品稀释液稀释一定倍数（n倍）后再测定，计算时请Z后乘以总稀释倍数（×n×5）。5．封板膜只限一次性使用，以避免交叉污染。6．底物请避光保存。7．严格按照说明书的操作进行，试验结果判定必须以酶标仪读数为准.8．所有样品，洗涤液和各种废弃物都应按传染物处理。9．本试剂不同批号组分不得混用。10.如与英文说明书有异，以英文说明书为准。保存条件及有效期1．试剂盒保存：2-8℃。2．有效期：6个月[详细]
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产品样册
半乳糖凝集素9蛋白galectin 9 peptide说明书 DESCRIPTIONSourceE.coliderivedAla2Thr323Accession#BAA31542NterminalSequenceAnalysisAla2Structure/FormMonomerPredictedMolecularMass35.8kDaSPECIFICATIONSSDSPAGE34kDa,reducingconditionsActivityMeasuredbyitsabilitytoinduceapoptosisofJurkathumanacuteTcellleukemiacells.Lu,L.H.etal.（2007）J.Biochem.141:157.Theforthiseffectistypically15ug/mL.Measuredbyitsabilitytoagglutinatehumanredbloodcells.Hadari,Y.R.etal.（2000）J.CellSci.113:2385.Theforthiseffectistypically2.5-12.5μg/mL.EndotoxinLevel<1.0EUper1μgoftheproteinbytheLALmethod.Purity>95%,bySDSPAGEunderreducingconditionsandvisualizedbysilverstain.FormulationLyophilizedfroma0.2μmfilteredsolutioninMOPS,NaCl,EDTA,DTTandTrehalose.SeeCertificateofAnalysisfordetails.PREPARATIONANDSTORAGEReconstitutionReconstituteat100μg/mLinwater.ShippingTheproductisshippedatambienttemperature.Uponreceipt,storeitimmediatelyatthetemperaturerecommendedbelow.Stability&StorageUseamanualdefrostfreezerandavoidrepeatedfreezethawcycles.l12monthsfromdateofreceipt,20to70°Cassupplied.l1month,2to8°Cundersterileconditionsafterreconstitution.l3months,20to70°Cundersterileconditionsafterreconstitution.BACKGROUNDGalectinscompriseafamilyofmultifunctionalcarbohydratebindingproteinswithspecificityforN-acetyllactosaminecontainingglycoproteins.Atleast14mammalianGalectinssharestructuralsimilaritiesintheircarbohydraterecognitiondomains（CRD）,formingthreegroups:prototype（oneCRD）,tandemrepeat（twoCRDs）,andchimeric（oneCRD,uniqueN-terminus）（1,2）.FulllengthGalectin9isawidelyexpressed39kDatandemrepeatGalectinthatcontainstwoCRDsconnectedbyalinkerregion（3）.Progressivedeletionwithinthelinkerregiongeneratesa36kDaisoform,alsoknownasEcalectinorUAT,aswellasa35kDaisoform（4）.ThisrecombinantproteincorrespondstotheEcalectinisoformofhumanGalectin9andshares70%and73%aasequenceidentitywiththecorrespondingregionsofmouseandratGalectin9,respectively.Galectin9exhibitsawiderangeofactivities.Allthreeisoformsfunctionaseosinophilchemoattractants（5,6）.Thisactivityisdestroyedbythrombinmediatedcleavagewithinthelinkerregionofthelongisoform,althoughtheEcalectinisoformisresistanttothrombin（7）.Galectin9bindstocarbohydratemoietiesofIgE,therebypreventingimmunecomplexformation,mastcelldegranulation,andasthmaticandcutaneousanaphylaxisreactions（8）.Independentofitslectinproperties,Galectin9inducesthematurationofdendriticcellswhichpromoteTh1polarization（9）.Galectin9inducescellularapoptosisinpartbydirectbindingtoTIM3（10,11）.ItsinteractionwithTIM3inhibitsTh1cellandCD8+cytotoxicTcellresponsesandalsopromotesregulatoryTcelldifferentiationandactivity（11,12）.Galectin9suppressestumorcellmetastasisbyinterferingwiththeassociationsbetweenhyaluronicacidandCD44andbetweenVCAM1andIntegrinα4β1（13）.TheEcalectinisoform（UATuratetransporter）canalsobeexpressedasanintegralmembraneproteinandmediatethecellulareffluxofurate（14）.References:1.Yang,RY.etal.（2008）ExpertRev.Mol.Med.10:e17.2.Elola,M.T.etal.（2007）Cell.Mol.LifeSci.64:1679.3.Tureci,O.etal.（1997）J.Biol.Chem.272:6416.4.Chabot,S.etal.（2002）Glycobiology12:111.5.Matsumoto,R.etal.（2002）J.Immunol.168:1961.6.Sato,M.etal.（2002）Glycobiology12:191.7.Nishi,N.etal.（2006）Glycobiology16:15C.8.Niki,T.etal.（2009）J.Biol.Chem.284:32344.9.Dai,S.Y.etal.（2005）J.Immunol.175:2974.10.Seki,M.etal.（2007）ArthritisRheum.56:3968.11.Zhu,C.etal.（2005）Nat.Immunol.6:1245.12.Sehrawat,S.etal.（2010）PloSPathogens6:e1000882.13.Nobumoto,A.etal.（2008）Glycobiology18:735.14.LealPinto,E.etal.（2002）Am.J.Physiol.RenalPhysiol.283:F150.[详细]
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Optimization of LC-MS peptide mapping
Peptide substrate identification for yeast Hsp40 Ydj
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