[ - 1 - ] cell apoptosis detection BMS306FI rh Annexin V FITC KIT 300 test 5000 ANNEXIN V rh Annexin V FITC KIT 200 test 4880 BMS306FI/100T rh Annexin V FITC KIT 100 test 2850 BMS306FI/c rh Annexin V FITC KIT 020 test 1300 8008282330 8008571184
[ - 2 - ] -1 1 8008282330 8008571184
[ - 3 - ] 8008282330 8008571184 1
[ - 4 - ] 1 1 10 6 2 PBS 200g 5 3-20 C 1ml 4 30min 4 C 5200g 5 6 2 4 1ml PBS 8008282330 8008571184
[ - 5 - ] Hoechst 8008282330 8008571184
[ - 6 - ] eochst 33342 / PI 8008282330 8008571184
[ - 7 - ] 8008282330 8008571184
[ - 8 - ] 8008282330 8008571184
[ - 9 - ] 7-8 8008282330 8008571184
[ - 10 - ] -9 8008282330 8008571184
[ - 11 - ] PI COULTER BD 2 1 PBS 5 10 6 1500rpm 400l 1 Binding Buffer 2 abcde 1 10 6 3a 4b 2% 30 Annexin V PI 5c 10l PI 15 6d 5l Annexin-V 15 7e 10l PI 5l Annexin-V 15min 8 400l 1 Binding Buffer 3 1 a Annexin V vs PI R1 2 d FITC FL2 3 3 c PI FL1 4 b 90% 5 e 8008282330 8008571184
[ - 12 - ] 8008282330 8008571184
[ - 13 - ] DNA - 12 8008282330 8008571184
[ - 14 - ] 12 8008282330 8008571184
[ - 15 - ] T DEX DNA SSC 14 8008282330 8008571184
[ - 16 - ] 8008282330 8008571184
[ - 17 - ] 8008282330 8008571184
[ - 18 - ] ISEL 16 8008282330 8008571184
[ - 19 - ] Dig-11-dUTP DNA 3 -OH 1% Fc FC [ ] [ ] 1 PBSpH7.4 PBS50mmol/LNaCl 200mmol/L 2 K200g/ml K 0.02gPBS 100mlpH7.4 3 2% 30%2.0ml 98.0mlpH7.4 4TdT Trizma 3.63g 0.1N HCl ph 7.2 1000 [CH 3 2AsO 2 Na. 3H 2 O] CoCCl. 2 6H 2 O 5TdT TdT 76lTdT Dig-11-dUTP 6 NaCl 17.4g 8.82g 1000ml 70.05%W:V DAB DAB 5mgPBS 5mlph7.4 H 2 O 2 0.02%V:V 80.5%W:V 0.5g0.1mmol/L 100mlpH4.0 9100% 100%95%90%80%70% 10% 10 ONCOR 8008282330 8008571184
[ - 20 - ] [ ] 1 5 3 95%75% 3 PBS 5 K 20l/ml 5 4 2 2 10% PBS 5 2:1-20 5 PBS 5 2 510 7 4% 10 50-100l PBS 5 2 2 2% PBS 5 PBS 5 3 2 TdT 15 4 54lTdT 37 1 TdT 5 37 30 10 6 PBS 3 5 30 7 PBS 5 8 0.05%DAB 36 9 4 3 1 1 5 10 3 10 30 100% 3 11 3 2 DNase 1mol/L 34 TdT 2 dutp/ 8008282330 8008571184
[ - 21 - ] Dig-11-dTUP TdT DNA 3 -OH 494nm 525nm DNA DNA DNA 10 38 10 [ ] [ ] 1 PBSpH7.4 50mmol/LNaCl 200mmol/L 2 K200 K0.02gPBS 100mlpH7.4 3 1% PBSpH7.4 4100%95%90%80%70% RNA 5Trizma ph7.2 Trizma 3.63g 0.1N HCl ph 7.2 [CH 3 2AsO 2 Na. 3H 2 O] 29.96g CoCCl. 2 6H 2 O 0.238g 1000ml 6TdT 76lTdT 32l 7 NaCl 17.4g 8.82g 1000ml 8 FITC 1%BSA PBS 56l ONCOR49l 9 0.1% Triton X-100 PBSTriton X-100 1.3mlPBS 128.7mlpH7.4 10 PI PI 50g RNA 5000 0.5mg PBS 10mlpH7.4 [ ] 1 1 5 3 95%75% 3 PBS 5 K 20 15 4 2 2 2 10% PBS 5 21-20 PBS 5 2 3 510 7 /ml 4% 10 8008282330 8008571184
[ - 22 - ] 50100 1 1 2 2 1 7 20 2 8008282330 8008571184
[ - 23 - ] 3 510 7 /ml 4% 10 50100µl PBS 5 2 23%H 2 O 2 30min 30.15mol/L PBS 2 5 0.15mol/L PBS 2 5 K 020 70.15mol/L PBS 2 5 8TdT 10min 9TdT 37 1h 10 0.15mol/L PBS 2 5 dutp - 17 8008282330 8008571184
[ - 24 - ] [ ] [ ] 1 PBSpH7.4 2 K200g/ml K 0.02gPBS 100mlpH7.4 3100%95%90%80%70% 10% RNA 4TdT ph7.2 5TdT 76µlTdT 32µlDig-11-dUTP 6 µl ONCOR49µl 8 0.1% Triton X-100 PBSTriton X-100 1.3mlPBS 128.7mlpH 7.4 4 9PI PI50µgRNA 50 000 0.5mgPBS 10mlpH7.4 1 1210 6 0.5ml PBS 1% PBS 15 4 1000rpm 5 10ml PBS `5 70% -20 5 2 41000rpm 5 1ml PBS 1.5ml 3 2 TdT 50µl TdT 37 30 15 4 1.0ml 1.0ml 5 100µl 30 15 6 1% Trito X-100 PBS 7 1.0ml 15 8 17 510550nm 620nm PI DNase I 1µM34 8008282330 8008571184
[ - 25 - ] TdT µl TdT 100mmol/L DTT 0.1mmol/L CoCl5mmol/L BSA0.05mg/ml TdT 20UBiotin-dUTP 1nmoles 2 4SSC1SSC=0.15mol/L 0.015mol/L Triton-X 100 0.1% 5%w/vAvidin-FITC 2.5µg/ml 8008282330 8008571184
[ - 26 - ] 8008282330 8008571184
[ - 27 - ] 8008282330 8008571184
[ - 28 - ] ISNT ISNT DNA 3 -OH 5 Klenow 5-3 3 -OH 5-3 DAB DNA 8008282330 8008571184
[ - 29 - ] 1 2 2SSC 80 20min 30.5% ph 2.037 020minPBS 4 Buffer A 5min 5 Buffer A 50 2SSC 80 20min DNA I Klenow 8008282330 8008571184
[ - 30 - ] 8008282330 8008571184
[ - 31 - ] DNA density gradient centrifugation pulse field gel electrophoresispfge field inversion gel electrophoresisfige DNA high-molecular weight DNAHMW DNA 50~300kb HMW 180~200bp DNAlow-molecular weight DNA LMW DNA 21 8008282330 8008571184
[ - 32 - ] 8008282330 8008571184
[ - 33 - ] 2RNase TE 10mg/ml10015min DNase 8008282330 8008571184
[ - 34 - ] DNA DNA DNA %= DNA / DNA+ 1 10mmol/L Tries1mmol/L EDTA0.2%Triton X-100pH7.5 2 TAE0.001mol/L EDTApH8.00.04mol/L Triton- 1 1 10 6 10000r/min 5min 2 0.4ml 1h 313000r/min 10min Eppendorf 50% 0.5mol/L NaCl 5min 412000r/min 10min 5 TE100l65 10min 20l 12l 1 PC ph7.8 0.2mol/L Na 2 HPO 4 192µl 0.1mol/L citric acid8µl 2 K 1mg/ml 8008282330 8008571184
[ - 35 - ] DNA 50300kd DNA DNA DNADNA 50 300kb DNA DNA DNA DNA 1 2 3 4 5 5 000kd DNA [ ] 1PBSpH7.4: K 2 HPO4 1.392gNaH 2 PO4. H 2 O 0.276gNaCl 8.770g 900ml 0.01N KOH ph 7.4 1000ml 2 0.1mol/L EDTApH 8.00.01mol/L Tris-HClpH7.60.02mmol/L NaCl 310mg/ml K 410% SDS 54mg/ml PMSF4mg PMSF 1ml -20 8008282330 8008571184
[ - 36 - ] 6TE 10mmol/L Tris-HClpH7.61mmol/L EDTA 710mg/ml [ ] 1 PBS 5 10 7 3 2 1% 42 3 4 50100l 5 1cm 2 45l1.5 10 6 510g DNA 6 50 1mg/ml K 1% SDS 50 24 50 4 7 50 40 g/ml PMSF TEpH7.6 50 1 8 65 DNA DNA DNA G 1 Hotz ph7.8 / DNA DNA DNA ladder PI DNA ladder [ ] 1 2 [ ] 1 / ph7.8: 0.2mol/L Na 2 HPO4 A0.1mol/L B 192ml A 8ml B ph 7.8 20.25% v/vnp40:0.25ml NP 40 100ml 31mg/ml RNase A:1mg DNase RNase 1ml 41mg/ml K 5DNA 0.25g 70ml 30ml 8008282330 8008571184
[ - 37 - ] 6DNA 100bp DNA ladder 7 10 TBE 54g Tris 27.5g 20ml 0.5mol/L EDTA ph8.0 8 EB11mg/ml [ ] 14 10 6 10 7 10ml 70% -20 4-20 2 1000g 5 40l / 0.5ml Eppendorf 30 3 1500g 5 Eppendorf 15 4 3l 0.25% NP40 3l RNase 37 30 5 3l K 37 30 6 12l 0.8% DNA 0.5 TBE 2V/cm 1620 7 50100l PBS PI FCM DNA ladder 1 2 3 4 DNA 5 DNA DNA DNA 6 DNA DNA ELISA 22 POD DNA DNA POD DAB POD 8008282330 8008571184
[ - 38 - ] DNA DNA 22 [ ] 96 24 [ ] 1 2 1ml POD-DNA 1ml 3 0.1mmol/L ph9.2 13.6g 7.35gH 2 O 950ml 1 N HCl 1N NaOH ph 9.2 1000ml 4 ph9.2 5 1l 9l 0.1mol/l PBS ph7.310 NaCl 80gKCl 2gNa 2 HPO 4 7H2O 11.5gKH 2 PO 4 2g 1000ml 1 10 1:10 137mmol/L NaCl2.7mmol/L KCl 4.3mmol/L Na 2 HPO 4 7H2O1.4mmol/L KH 2 PO 4 6 0.05%NaN 3 PBSNaN 3 0.5g 1000ml PBS 7 0.05% Tween-201mmol/L EDTA0.25% BSA 0.05%NaN 3. 8 1l POD-DNA 9l 9 5075mg DAB 100ml 0.05mol/L Tris-HClpH7.4 4 0.0100.003% H 2 O 2 10 124 4 8008282330 8008571184
[ - 39 - ] 4 2 3 4 5 6 7 8 9 [ ] 3 enrichment factor EF ( ) EF = ( ) [ ] ELISA 15 1/10 3%8% ELISA 5 5 10 4 1500rpm 5 5 10mmol/L Tris-HClpH7.4400mmol/L NaCl5mmol/L CaCl 210mmol/LMgCl 2 37 2 1500rpm 10 400 8008282330 8008571184
[ - 40 - ] 1:5 HL-60/CAM ELISA 5 10 2 /ml 50 8008282330 8008571184
[ - 41 - ] 8008282330 8008571184
[ - 42 - ] 8008282330 8008571184
[ - 43 - ] V = V 1 1 V 1 (min µ g ) = ( ) VDNA( µ g) 8008282330 8008571184
[ - 44 - ] 8008282330 8008571184