血小板の活性化、染色と解析 概要 必要な試薬と装置
Contents
1. ebi 0 196 PBS edel cx Jp sep dk D2. ACD BD G64816 2 FALCON 125x75mm BD 352058 3 4 ADP 2x10 4M Bio Data Inc Phila PA 101312 5 0 19 PBS 196 8C 1
3. g PAC 10 L RGDS 1 RHGDS PAC 1 PAC 1 10 5 5 50 L
4. BD ACD EDTA in Vo PAO 1 pH CaF PAOC 1 EDTA PAC 1 ACD 3 S0 2mL
5. 3 6 5 SAER WBC 39 G 5 15 0 1m 5 FL3 Threshold FACO X m9 e FACS COM TZ KT EE CaliBRITE MARITI Lyse No Wash l FSC SSC
6. 64 O FACS in VO
7. ACD 10 34 65 BD ADP Phorbol1 5 193 PMA TRAP 1 10 ADP 50 L 1 75mm 2 0 45mL 3 GO0 25C 4 PAC
8. 3 FLG 5 4 FL 1 FL PMT ud occ LLS0 LIAR SONWNS Mew 10 PAC 1 PAC 1 HGDS 17 o FL3 d PAC 1 FITC IgG PE CD61 PerCP FLG FITC PAC 1 FITC FL 1 PE RGDS PAC 1 FITC CD62P PE CD61 PerCP PAC 1
9. CD62F 1 10 100 L 2 80 196 PBS 1mL 1 5mm HA 100 L 20 1 e 2 8C 5 8C 3 1 200xg 5 20 25C gud 4 1 mL 5 1 200 x g 5
10. platelets and WBCs 1 microparticles LL 1L LLL LL L1 LLLI LL LL LLLI LTI LLLLLU T TT rrr T TT DERE II EEEN Epub 100 101 10 103 104 FSC 6 H1 Yr E Lp LL 1 gp 1 iru LL LL LLUb LL Lb LLLI mu TI IITIN 100 101 102 103 104 PAC 1 FITC 8 ADP H1 71
11. pr 6 1 mL CD6 1 CD42a Threshold CD62P PAC 1 CDG 1 CD62P PAC 1 6 3 l 2 O 1 5x 5mm
12. 6 5 000 10 000 7 WBC 8 Jackson AL Warner NL Preparation staining and analysis by flow cytometry of peripheral blood leukocytes In Rose NR Friedman H Fahey JL eds Manual of Clinical Laboratory Immunology 3rd ed X Washington DC American Society for Microbiology 1986 226 235 Shattil SJ Motulsky HJ Insel PA Flaherty L Brass LE Expression of fibrinogen receptors during activation and subsequent desensitization of human platelets by epinephrine Blood 1986 68 1224 1231 Shattil SJ Cunningham M Hoxie JA Detection of activated platelets in whole blood using activation dependent monoclonal antibodies and flow cytometry Blood 1987 70 307 315 Abrams C Shattil SJ Immunological detection of activated platelets in clinical disorders Thrombosis and Haemostasis 1991 65 467 473 Abrams CS Ellison N Budzinski AZ Shattil SJ Direct detection of activated platelets and platelet derived microparticles in humans Blood 1990 75 128 138 Jennings LK Ashmun RA Wang WF Dockter ME Analysis of human platelet glycoproteins IIb IIIa and Glanzmann s thrombasthenia in whole blood by flow cytometry Blood 1986
13. 15 0 2 80 196 PBS 1 mL MR BRELES AE 8C 30 24 6Z 68 1 4 Bl5 EB8 FSC SSC FSC hreshold HT 7 4727 FL3 SSC
14. 68 173 179 69 70 SSC Mouse IgG PE 104 J WBCs and RBCs 4 103 2 tix 10 mr 101 ES E R2 debris 100 TT TT TT 100 101 102 103 10 FSC y xl PASS z 1 104 103 10 101 100 EL 1 T Hm 44 TT I 100 101 102 103 10 PAC 1 FITC RGDS 3 R px AE Counts CD62P PE 100 80 60 40 20 104 103 100 Mouse IgG CD61 PerCP PerCP 100 101 102 10 104 CD61 PerCP Ele RoO LL LL LUI LL LLLUL L pL uL Lud LTI LLLLIU TT T ITTTIH 100 101 102 103 104 PAC 1 FITC 4 ADP Re SSC Mouse IgG PE 104 ce x pug T3 ul i N 2 I LL Lu L 1 IIIll SSC R1 cS 4er Th Et EE 100 101 102 103 104 CD61 PerCP i ce ce 5 102 CD62P PE 10 100 TTTT rrr T TT E LITTITTIIH 100 10 10 103 104 PAC 1 FITC RGDS E 7 H1 TA i N 2 i ce TA i co i N 100
15. ulbecco s PBS pH7 2 0 2 PBS 0 2 m 6 8C bs wA 0 196 9 FBS PBS 8C BD Arg Gly AsD Ser RGDS Sigma A9041 PBS 10mg mL RHGDS PAC 1 Bx LPS FACS CaliBRITE CaliBRIT FACSComp COellQuest 3
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