活性化リンパ球の細胞内サイトカインの検出 装置 細胞
Contents
1. 10 ug mL BFA Fastlmmune KLH CD69 CD692. 1 BFA PMA I BFA CDD69 CD69 BFA 1000 SSC H 2 CD69 PE CD3 PerCP BD 340368 3 FL FL3 CDG3 CD69g CD69 90 1 Unstimulated Counts 109 CD3 PerCP 104 109 CD69 PE 10 1 CD69
3. CPT PBMC CPT 4 PBMC PBMC Ficoll Paque 109 FBS RPMI 1640 BioWhittaker 15 167F x 106 mL T X 10 mL FBS 53 54 PBMC 1xFACS Lysing Solution PBMC
4. BD 1 FACS Lysing Solution FACS Lysing Solution 10 PBS 3 FACS Permeabilizing Solution FACS Permeabilizing Solution 10 10 PBS 4 BD BFA 3 BFA
5. 9096 CD69 1 BFA PMA Ionomycin CD69 PE CD3 PerCP BD 340368 3 FL3 CD3d CD6g CD69 9096 2 5Z 58 1000 SSC H 100 80 Unstimulated Counts CD3 PerCP 104 100 CD69 PE 104 3 BD 20 L 12x75mm PMA 100 L 50 L
6. 1 12x75mm BD 3586058 5 796C0as 37 3 4 6 FACS BD 367673 Fastlmmune EDTA ACD 8 8 PBMC CPT BD 362753 PBMC CPT
7. PBMC RPMI PMA I x 108 HPMI 15 1 XFACS Lysing Solution mL 10 PBMC FACS Lysing Solution PMA 500xg 5 1 XFACS PermeabDilizing Solution 500 L 10 3mL 500xg 5
8. SEB a 10 g mL BFA 10 g mL SEB b 4 6 37C 3 CD CD2R BD 340366 a BFA 1 mL 820 L CD2 CD2H b 4 6 37C 55 4 CD3 a BFA CD3 b 4 6 37C CD3 CD3 FL3 CD5PerCP BD CD45PerCP BD 347464 CD3 BD 5 CD28 1 10ug mL CD28 BD 348040 SEB CD3 CD2 CD2R 56 Vo
9. 12X75mm BD 352058 1 PMA lonomycin PMA I PBMC RHPMI 1640 1 1 PMA ox 108 mL RPMI b 10 g mL BFA BFA 1 mL 20 L 25ng mL PMA PMA 1 mL 25 L 1 g mL Ionomycin lonormycin 1 mL 20 L c 37C 5 796CO CO 4 pH COs
10. 30 lt 3mL 500 x g 5 500 L 196 PBS S24 FACS5 CaliBRITE 1 1 FACSComp 93 0 2 AutoCOMP PMT FACSComp Lyse No Wash FastImrmune FACSComp AutOoCOMP FACSComp AutoCOMP BD
11. lonomycin 1 g mL lonomycin Staphylococcal enterotoxin B SEB Sigma S 4881 a PBGS 0 5mg mL b 4C C SEB 10g mL SEB Brefeldin A BFA Sigma B 7651 DMSO0 5mg mL b 20 L 20C C PBS 1 10 d BFA 10 g mL 245 BFA RPMI 1640 BioWhittaker 12 167F PBS DMSO Sigma D 8779 0 596BSA 0 196NaN PBS 4 196 PBS 4
12. FSC CellQuest LYSYSI FL3 2 CellQuest LYSYS II Attractors PMA FLS3 FSC SSC CD4 FSC SSC AS AIC US U10
13. PBS 196 BSA 109 DMS0 PBS 70C 3mL 5 500 xg FACS Permeabilizing Solution BD BD 1 OO0SOQ Phorbol 12 myristate 13 acetate PMA Sigma PB8139 DMS0 0 1mg mL b 20 L 60C PBS PMA 1 100 PMA 5ng mL PMA lonomycin Sigma 0634 0 5mg mL b 0C C PBS 1 10 d
14. AS AT1C US U 10C 59 60 Unstimulated 10 PMA lonomycin SEB o Hu IL 2 PE 102 103 10 co Hu IL 2 PE o Hu IL 2 PE iF 109 10 102 10 CD8 FITC a 72 2 1 27 8 FF T 104 CD8 FITC CD8 FITC 3 CD8TT FastlImmune CD8 T G 6 PMA IOnOmycin Staphylococcal enterotoxin B SEB PMA I CD8 CD8 CD4 IL 2 SEB IL 2 CD8 I Picker
15. LJ Singh MK Zdraveski Z et al Direct demonstration of cytokine synthesis heterogeneity among human memory effector T cells by flow cytometry Blood 1995 86 1408 1419 2 Openshaw P Murphy EE Hosken NA et al Heterogeneity of intracellular cytokine synthesis at the single cell level in polarized T helper 1 and T helper 2 populations JExp Med 1995 182 1357 1367 3 Ferrick DA Schrenze 1 MD Mulvania T Hsieh B Ferlin WG Lepper H Differential production of interferon g and interleukin 4 in response to TH1 and TH2 stimulating pathogens by g d T cells in vivo Nature 1995 373 255 257 4 Cherwinski HM Semenuk GT Ransom JT Stimulation of a T helper cell class 2 clone with immobilized anti T cell receptor antibody activates a Ca2 and protein kinase C independent lethal signal pathway JImmunol 1992 148 2996 3003
Download Pdf Manuals
Related Search