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Bpolyclonalantibody1975KohlerMilsteinBmonoclonalantibodyKohlerMilstein1984197587KohlerMilsteinContinuousculturesoffusedcellssecretingantibodyofpredefinedspecificity-hypoxanthineguanosinephosphoribosyltransferaseHGPRThypoxanthineHaminopterinAthymidineTHATHGPRT+HGPRT-HGPRT-HATHGPRTHATPEGSP2/0-Ag14X63-Ag8.653NSO/1MOPC-21CO2-704000r/min37100ml50ml25ml10ml1ml500ml250ml100ml10ml5ml1ml962450ml~50ul~200ul~1000ul2006-11(1)(2)BALB/cLOU/c8-12(3)B6-13422IgMIgG336-1106-1071-10ug2-42-410-100ug2-42-4A.20-400ug2-42-32-3B.10-50ug200-400ug24C.10-100ug24101-2()4-8BALB/c2-310%0.5-5ug/ml105-106/mlBALB/c376%CO23-521aRPMI-1640DMEMDulbercoModifiedEaglesMedium0.22um4•RPMI-1640RPMI-164096ml100L.G.1ml1ml7.5%NaHCO31-2mlHEPES1ml•DMEMDMEM13.37g980mlNaHCO33.7g10ml100L.G.10ml1NHClPH7.2-7.44•RPMI-1640DMEMRPMI-1640DMEM80ml15-20mlSP2/0•HTRPMI-1640DMEM99mlHT1ml•HATRPMI-1640DMEM98mlHT1mlA1mlbA100410-5mol/L:1.76mgAminopterinMW440.490ml1mol/LNaOH0.5ml100ml2ml/-20cHT(100,H:10-2mol/LT1.610-3mol/L):136.1mg(Hypoxanthine,MW136.1)38.8mg(Thymidine,MW242.2)100ml45-502ml/-2037dL-L.G.1000.2mol/L:2.92gL-L-glutamineMW146.15100ml4-5ml/-20e100:G1001g100ml4-5ml/-20f7.5%NaHCO3:NaHCO37.5g100ml4-5ml/4gHEPES1mol/L:23.83gHEPESN-2-Hydroxyethylpiperazine-N-2-ethanesulfonicacidN-2--N-2-MW238.3100ml4-5ml/4h8-100:200mg8-8-azaguanineMW152.14mol/LNaOH1ml99ml-201%20ug/mli50%PEG:PEG1000400020-50g60-800.6ml815-207.5%NaHCO3PH8.0SigmaGibco1210%65ml10114-16a48-36962-3100mlb50mlc1000r/min5-10d30ml10mle0.4%0.1ml0.9ml=4105/4=5106/2BALB/c75%510ml10ml1ml2001000r/min5-101-210ml1108-2.51085108-10108Feedercells10ml11000r/min5-105mlHATHAT2105/ml962104241053-5106961-2960.1ml2376%CO2A11082107-5107SP2/0-Ag1450ml30mlB1000r/min5-10C;40D1ml1454050%PEGPH8.01mlE10ml9020-30ml3720-3710F1000r/min5G5mlHATHAT80-100mlH960.10-0.15ml241.0-1.5ml376%CO2I5HAT1/2J7-10HTHAT14L1/1032100%/1011001AAAa.0.2mol/LNa2CO38ml0.2mol/LNaHCO317ml75mlPH9.6Tris-HClPH8.00.02mol/L0.1mol/LTris100ml0.1mol/LHCl58.4ml1000mlb.PH7.2PBSKH2PO40.2gKCl0.2gNa2HPO4•12H2O2.9gNaCl8.0gTween-200.5ml1000mlc.BSA0.1g100ml5-10%d.2mol/LH2SO4178.3ml98%21.7mle.PH5.0-0.2mol/LNa2HPO425.7ml0.1ml/L24.3ml50mlf.OPD490nmODOPD5mg10ml3%H2O20.15mlTMBSTMB450nmODTMBSTMB1mg/ml1.0ml10ml1%H2O225ulABTS410nmODABTS0.5mg1ml3%H2O22ulg.h.i.SPAj.-20-Na2HPO4100mgKH2PO4500mg150ml225ml125ml-1;1-20k.4096l.m.BELISAa.1-20ug/mlb.50-100ul/4372c.33-5d.200ul4372e.3-204f.50-100ul371-2g.50-100ul371-2h.50-100ul3710-30i.2mol/LH2SO4ODj.P/N2.1PN+3DCELISASa.PBS1108/mlb.100ul5%0.1mol/LNaHCO395ml+25%5ml372350ul/37-56200ul4372c.b50ul37-56-20153200ul4372d.3-204e.DELISAa.PBSb.1500rpm300.83%10c.100ulab51041500r/min15-114104-20d.EELISABALB/cIgMcAbELISAa.Ig100ul3724b.McAb371-2c.371-2d.371-2FABC-ELISAABC-ELISAELISABiotinAvidin4a.McAbELISAb.Ig100ul371c.100ul3730GDot-ELISADot-ELISADAB4-AgNO3HRPAPHRPAPa.2-5ul37b.3730c.2McAb3715HRPAPIg3730d.HMcAbIIPindirectimmunoperoxidaseAPAAPMcAbAa.b.PBSPH7.20.01mol/Lc.McAbd.e.FITCTRITCIgf.Ba.PBS410-2010ul1108/ml721mm-2010-15-20b.10-20ul370.5-1c.PBS15dIg10-20ul370.5-1e1591PBSf.FITCTRITCg.C4a.107/mlb.550mm100ul100ulMcAb430-90c.PBS900ml50ml4%NaN350ml1-5ml1000r/min5100ul430-90d.20-30ul10%10-100ug/mle.f.1%4PHAAa.Ob.PBSPH7.2c.NaN3d.Ba.50ml5PH7.2PBSNa2HPO4•12H2O3.58gKH2PO40.41gNaCl8.01g1000ml51500r/min5-1025%25%20%2.5137215PH7.2PBS42000r/min10PBS25%20%0.3%4b.LPS0.2mol/LPH8.0PB120ug/ml100137LPS6%37452000r/min100.01mol/LPH7.2PBS44%4c.McAbMcAb25ulV0.5-4%25ul303730-60Ca.O10PBS4-68%8%3%3%PBS17320%0.1%NaN34b.20%0.1ml1500r/min100.2mol/LPH4.0-1ml50ug/ml1ml453020PBS3-4PBS0.5-1%4c.McAbMcAba.1-100ug/ml100ul/4372b.100ul0.5-1%BSAPBS41-2c.BSA-PBS33d.100ul371-2e.125IIg100ul371-2f.-510342-3FACS1a.96b.HTc.d.a.b.20%HT2.515503c.5104-1105d.96320.2ml0.5310e.377.5%CO27-10f.g.bcd100.1ml12a.HTb.0.15mol/LNaCl2.0%2.0g100ml0.15mol/LNaCl1211510ml4c.d.e.45f.a.b.2.0%HT1%45c.1%3ml10d.1ml1%1mle.377.5%7-142mm24f.51CO20-201-20-402-19680%Aa.1-2b.10-20%5-10%DMSOHT0c.d.-70e.f.Ba.106-107/ml1mlb.c.-70d.2-4-70e.2-50375-10mlHT1000r/min10HT24377.5%CO2104-105/ml100%966A40SP2/062-68NS-154-640.075mol/LKCl-a.48-36b.100ug/ml-200.1-0.4ug/ml0.02-0.05ug/ml4-61000r/min10c.370.075mol/LKCl5ml3715-20d.311ml1000r/min10e.5ml20-301000r/min105ml4f.1000r/min50.5-1ml1-2g.10%Giemsa10-20GiemsaGiemsa0.5g33ml55-60233ml11/15mol/LPH6.8PBS910%Giemsah.100BIgMIgGIgEIgAAIgGIgELISAAMcAb10-20Ig10-20a.IgGIgG1IgG2aIgG2bIgG3IgMIgAb.0.06mol/L-PH8.6c.1%1g100ml0.06ml/LPH8.6-0.02%NaN34d.3mma.10mlPEG6000PVPpolyvinylpyrrolidone40.5-1ml-20b.1%3mlc.16d.IgG3712-244BELISAa.ELISAb.c.IgIgHRPIgd.a.100ulIg22b.200ul1c.2100ul4d.4100ulIg2e.4100ulHRPIg2f.g.HRPIgea.100ul/4b.100ul/371c.HRPIg100ul/37152mol/LH2SO4OD490CPAGESDS-PAGEIEFWBPAGESDS-PAGEIEFWBPHKELISAELISAELISAELISAELISA100ul/40.5%BSA-PBSPH7.2100ul/371410100ul/371HRPIgG100ul/371OPD100ul/37152mol/LH2SO4495nmAKA0/A0-A=1+K/a0A0=AA=Aa0=K=11BALB/cBALB/cIgSPFAa.BALB/cb.Pristanec.Ba.0.3-0.5mlb.7-10PBS5105/0.2mlc.5162-35-10mld.2000r/min5-702anchoragedependentcellADC10-15%1106-2106/ml10-50ug/mlA.B.MicrocarrierCytodexIBiosilonSuperbeadsC.-100-500um25-75umD.3DMEMRPMI-164010-20%21A.2000r/min15PH7.2VBS0.004mol/L0.15