BrdU Flow Kit - 日本ベクトン・ディッキンソン
Contents
1. 2 16 22 BrdU 10 mg mL DPBS OQ n vivo DOOD 100 phi mg MOOO000000 Das 0 8 5 7 HERETO B BrdU Flow Kit 0000000 1 a cells 50 uL Staining Buffer 6 FALCON cat 345002 00000 b 7 00000 Staining Buffen BD Pharmingen Stain Buffer TJ E UO UO B UOI UI D2. Pharmingen BrdU Flow Brdu Flow 3 FACSCalibur OOOOOO00000004000000 FACSCalibur 0 0 0000000000400000 1 2 4 40 D 24 Section 7 UU DEPT E E TEE EE BEABEDEEUERU THE PEt 20 6 51
3. HOO Lyse Washt i di B B CD D PMT Section 6 1 D U B D U D D BD Pharmingen BrdU Flow Kit 1 D LH HH H BrdU EF D HH E E H HL E E HL EE DB DE ET ELI METER TE 23 2 BD Pharmingen BrdU Flow
4. 13 14 10 O03 D10 G4 19 000 BrdUQG OOOO FIC anti BrdU 1 0 EH 0 DNATI OOD 7 AAD0000000000000030000D10 G4 10 0010 uM BrdU 7 vs Flow Kit OOOUOOUOO 000 Sub G0 G1pR6e0 0000000000560 M0000 1 R3938 69 MS R4038 60 WOU G2 Mj R50 14 Thymus 2 Hours BrdU FITC 100 150 200 2 50 100 150 200 250 0 50 100 150 200 250 7 AAD Bone Marrow A 40 Minutes B 2 Hours C 4 Hours 50 100 150 200 250 7 AAD 4 C57BL 60 000 Brdu 1 4 2 OHOU4000000000000000 0000
5. 4 2 MFis10I 0 D U U U D 2 HOU 2 Uu 19 20 FLI sf 2 ei FE 25i FL2 5403 1 es FL 1 FL3 Tae SE 3 oof FL P d P FL3 1 97 9 FL4 E i iia FL4 X3 FL3 E LJTaur tear SS Param falla EGG 1 42 Ein 7 M 32 ran 8 sz oo aaa E j OE peri Param TIT 8 anti IL LO FITC anti BrdU 0 7 AADI Hl B DB H 5D U D HU 7E HT PMT anti iL 100 0000000000 8 1 4 Parum Ur Falle M FLI 0 3 3 FL2 lt j 5 73 sz suc tool Us FL2 34 0 3 s r1 es anf 2 8 FL3 Tr XE Mm i 8 14 FH mx i FL3 10 8 FL2 5 ns E gp 5 FL3 4 977
6. 5 UU UD FITC anti BrdYj FLIM O 0 7 ADD 2 anti IL 10 OOO PE anti TNF oD 0000000000 5 FACSComp 0 00000000000 Eyse Washt D U U D CE UU 0 5 810 B20 BD CellQuest 0 0 0 D UD DD B B DU DU B D D U CD UD I 000 D 0 00 CD 0 2 1 volager 000000000000 DDDUFSC vs SSCDDDBBDIBULULUS AITITI BD UO B B 7 U UU CE UU C 5 B21T 5 5 2 7 5 3 FL3PMT voltage G0 G1 II H 3B HU TH HU D Hl D H ED MFE 50000 2567 2 2 10 17 18 FLI ipe Fz THE mz g I FL2 E FL1 n Fi a Fi2 O0 0 s FL3 um 2
7. EE 11 BD Pharmingen BrdU Flow Kit L L H D C H D EL EE EL D E 0000 BrdU BrdU IL 10 O00 AH 7 AAD vs FITC anti BrdU 5 HOOOOOUOUUU0O Quadrant Marker rn 10 Dd d 2700 i B APC anti CD8 vs FITC anti BrdU 000 c0 APC anti CD8 vs PE anti IL 10 0 340 0 L 100 000000000000 0 00 D PE anti IL 10 vs FITC anti BrdU HOOUOUUBrdUA OOOOUUI IOQQOOUU0D0U038000000 1 EE BALB c 00000000 n vitrot t C HD C DD B 7 D UU 7 CI CE CEI UU L 000045000000210 FITC anti BrdY FLI T PE anti IL 10 FL2 T 7 AA D APC anti CD 4
8. tu in vito 0 00000 BrdU 10 plo i1 mM BrdU 1 5 OO0 n 10 yuh 1 mM BrdU 1x DPBSIIT I HAU 5 cCTLL 20 00000 42 EE E E EE E
9. 0 MOL 3 7 1 8 0 1 DNasd TOU00000 300 LU DNase 1 mg m 1xDPBST I D00 00001000 5 7 LO 1xDPBSL 300 ug mL Working Stock Solution L 00 HOO 5 6 30 pl DNase 1 mg mL 0 000000 1 mg mL DNase j 80 QIO D EB LU D U DNase d mg ul Working Stock Solution DNase 30 105 cells OO 037 Q OOOOUO000PNasef 5 0000000000 0 7 AAD 7 amino actinomycin 7 1 068 cels samplert 0000000020 LO 7 AADQ OOO 4 HUETE HE Staining Buffet 1xDPBSO 3D D UO DU U 000 D D 0 m 0 09 w w TI 0 000000 HOO guguduuguuuuut BD Pharmingen Stain Buffet FBS Cat No 554656000000 1x DPBS 0 T 1 000 KCI 0 2g K HPO 0 2g NaCl 8 0g Na HPO 7H O 2 16g pH7 2 7 4 Section 3 BrdU 0 00 000 BrdU Flow Kit 0 000000
10. 2 d 00000001 Staining 2 5 2 Cytofix Cytoperm Buffer Cat No 2090KE JOU QO000000000 00 8 uly Cytofix Cytoperm Buffer D B 0 BL UL EE BE BE OE D 00 15 30 c 1mL Perm Wash 3 000 Cytoperm Plus Buffer 2356 a 000000100 LO Cytoperm Plus Buffer 000000000000 c 1 5 100 0 O 4 000000 a 00000100 uly Cytofix Cytoperm Buffer D B 0000000000 5 c 1 mL Perm Wash 1 0 5 DNas j Cat No 2358KC II 0000000000000 srdutg ttu ttu tltl ue2 8 5 300 pg mL 30 ug DNase tube TIl p OO0100037 Q 0000000000000 c 1 mL Perm Wash 6 BrdU 1 a b B B 7 L7 C U UU 8 835 Buffer 50 uL Hu UBL Lu 2 c 1 mL
11. E EEEE E a ENE EEE EE E EEEE ENG EEE EE E EGE E EE E 12 CD8 PE cess T T CD4 FITC CD4 FITC 2 BrdU Flow anti CD A4 PE anti CD 8 O APC anti B220ITI 0 0 0 UCyofix Cytoperm Buffer DU LI Pharmingen 2 5 CDS PEM LI 2 2 APCOvs CDS8J PE B D E CDA CDS I 0 B220 I 0 2 2 Positive MFI Conventional BrdU Conventional BrdU Protocol Protocol Protocol Protocol CDA1 FITCH 20 5 2345 278 141 CD8f PE 8 4 9 0 649 195 B2201f APC 62 9 60 1 261 189 02 020000 2
12. O EE EE BERGE BOE BEI Section 1 BD Pharmingen BrdU Flow Kit DO B0 D D U BD Pharmingen BrdU Flow Option l Option 2 E E a Pharmingen BrdU Flow Section 1 BrdU Flow 000000000
13. ERE E E ED EE BE T BILE Flow BrdU Flow Flow a EE B ELEEEEE D DEF EEEE DEI BEBE BEES BD Pharmingen BrdU Flow
14. PS ad FL3 FL2 TM E r3 row bo Lim 3 EL FL4 A liga d FL4 FLS T ree arts B L Frar rale EDH Farum FUT 4d 6 1 2 anti BrdU Y Tj FL2 2 Uu Bl fi Compensation 22 25 B2 Sern Parum Teig Hiis FLI 0 568 FL2 rer m rz inb 1 Im FL PA fii ami sue lg 5 p4 sus FL2 1 01 3 BS PT 15 FL3 FL Ph na noel Lm TT P rw Lin LJfaur d ginr Farem FLE D 7 7 p
15. OHO0001xPerm Wash Buffer 4 C O G 000000000000 200 10xPerm Wash Buffer 10 DU 4 Q O UO UU DE UU U OOO 10xPerm Wash 1xPerm Wash Buffer 0 45 um OOOUOOU0000 00000000 000000000000 Perm Wash Buffeg Cytoperm Plus Buffer Cytoperm Plus Buffer BrdU Flow Kit D E D D E 00 2 uL sample T Cytoperm Plus Buffer 10 Cytoperm Plus 5 mg mL BraU j 0 5 BrdU j j 0 22 n vitro 1D 8 in wo D 0D mg mL BrdU 0 31 1 mLO 1xDPBSQOOOOUOOO000U000000 3200001 mM mMO 010
16. 7AADD D HD D UO U 2 BrdU 5 BrdU positive 7 t D D OU 0 U U D U U 15 16 Section 5 ERE DE EE DE LED ABI EI LETTRE e TI zz te dE BB S EHE Ce 0 zer Hee ED BE HEEL BUE B TEE EI S at ee e ES E ui EAD E PB BIELLA acp Ewleidar AM FL1 uis 2 rac BRE 3 Us 13 TE fi ini FL2 2 8 FLI m jl inj Al i FL2 FL3 m I J TE saw i T T FL3 FL FLN LIA FLA w i au E Lin FL3 i m FL4 LI Tour Cahir Fara m TE 5 BrdU Flow Flow
17. BrdU in vitro in vivo BrdU 1 B uult Option 1 OOUU 4 0 Cytofix Cytoperm Buffer Dmm ai Staining Buffer uU D D Cytoperm Plus Buffer uU uut ETEPELHETELEHE ELNE Staining Buffer Uu Cytofix Cytoperm Buffer DL UL OL CL OE D U D E DNaseD D DU U BrdUr DD Bad d d trm 20 FITCO OO BrdU D H1 D 3 7 AD0 0000000000000 DNAQ 0 40 Staining Buffer 0000000 100 dimethyl sulfoxid amp FBS T 2 Perm Wash o ET Section 2 1 Component Storage condition of vials FITC anti BrdU Antibody 4 C 1 Cytofix CytoP erm Buffei D D LI 0
18. BrdU FLOW Kir Kit Manual BD Biosciences Clontech Discovery Labware Immunocytometry Systems Pharmingen amp BD Ee Co e FITC anti BrdU Antibody Kit Component 2361411 10000 e Cytofix Cytoperm Buffer Kit Component No 2090KE 1 e Perm Wash Buffer 10x Kit Component No 2091KE 2 0000 e Cytoperm Plus Buffer Kit Component No 2356KC 1 e 7 AAD Kit Component 2359KC 1 0000 e Kit Manual 80 e BrdU Kit Component No 2420KC 5 e DNase Kit Component No 2358KC 5 0000 BD Pharmingen BrdU FLOW KIT MANUAL UU Section Uu ul BL a TEM PECUNMTHREEERKPEREEEREEMM 1 1 0 BD Pharmingen BrdU 2 4 3 Brdu 7 Pe cse obtu tpe ctetu eb rt hians 7 B BrdU Flow Kit DH HD BD sss RR 9 A ac atl e ete m cms 11 EET isl edente dale atri ra dna 16 SNR ERN RR TENTER 23 Des Inn dl 25 te assis ort EE tenu a eae UMS LI IE UE 32
19. D bremodeoxyuriding 5 anti BrdUr D D D amino actinomycin p 7 2 7 0 1 A a SS BD Pharmingen O0 00O BrdU Flow Kit Lj 1 1 Hl D DB D H H BrdUC E B E HL D DE D ELT IEEE EDGE EL E E E
20. Perm Wash Buffer 00010000000000 100 0 0 O 7 00000 D D BE OE U DU C U D C DNAD HOO 8 8 7 AADQ 2 8 a 0000001 Staining 4 00 5 4 HOU 10 Section 4 ETT EDS DEP ET DIEI DISP EE E E RC E 488 isothiocyanate FITCO FL10 0 O phycoerythri FL2 00 0 O 7 AAD 5 55 5 rFAcsSCalibu 1 1 r1 1 O B E S geo CI ELE SP EEG EE E S ELE EE EPA E E 7
21. Specificity Clone Isotype New Cat No Old Cat No Format Size Human Cytokines and Chemokines 364 3B3 14 Mouse lgG1 554560 18934A FITC 0 1 mg 554561 18935A PE 0 1 mg IL 2 MQ1 17H12 4 RatlgG2a 554565 18954A FITC 0 1 mg 554566 18955A PE 0 1 mg 559334 18955X PE 100 Tests 554567 18959A APG 0 1 mg IL 3 BVD3 1F9 Rat IgG1 554676 20575A PE 0 1 mg IL 4 MP4 25D2 Rat IgG1 554484 18504A FITC 0 1 mg 554485 18505A PE 0 1 mg 554486 18509A APC 0 1 mg IL 4 8D4 8 Mouse IgG1 554516 18655A PE 0 1 mg 559333 18655X PE 100 Tests IL 5 TRFK5 Rat IgG1 554395 18055A PE 0 1 mg 559335 18055X PE 100 Tests 554396 18059A APC 0 1 mg IL 5 JES1 39D10 Rat IgG2a 554489 18515A PE 0 1 mg 559332 18515X PE 100 Tests IL 6 MQ2 13A5 Rat IgG1 554544 18874A FITC 0 1 mg 554545 18875A PE 0 1 mg IL 6 MQ2 6A3 Rat IgG2a 554696 20654A FITC 0 1 mg 554697 20655A PE 0 1 mg 559331 20655X PE 100 Tests 25 Specificity Clone Isotype New Cat No Old Cat No Human Cytokines and Chemokines Continued IL 8 IL 10 IL 10 IL 12 p40 p70 IL 12 p70 IL 13 IL 16 GM CSF GRO IFN y IFN y IP 10 MCP 1 MCP 3 MIG 26 G265 8 JES3 9D7 JES3 19F 1 C11 5 14 20C2 JES10 5A2 14 1 BVD2 21C11 10G4 B27 4S B3 6D4 D6 G2 5D3 F7 9H11 B8 11 Mouse IgG2b Rat IgG1 Rat IgG2a Mouse IgG1 Rat IgG1 Rat IgG1 Mouse lgG2a Rat IgG2a Mouse IgG1 Mouse IgG1 Mouse IgG1 Mouse lgG2a Mouse IgG1 Mouse IgG1 Mouse IgG1
22. 4 C 1 Perm Wash Buffer 10x 4 C 2 Cytoperm Plus Buffer 4 C 1 BrdU 10 mg ml 80 C 5 DNase 8 0 C 5 7 AAD 4 C 1 PB DPBSITI D Permn Wash FITC conjugated anti BrdU antibody 5 ulf FITCO O0 O BrduUg Og cells sample QO OO00000000 1xPerm Wash Buffer 0010500 n d guuuultttttltlliulululus59 uiui OHOO00 00 FiTC conjugated anti BrdU antibody 044 Q OO DL EE HE HE UE EE C ytofix C ytoperm Buffel 0 0 0 B Cytofix Cytoperm Buffer LH D E UO UO OL UL D ES ETE EC E ERE ET EAE E BE EET E 25 ml Cytofix Cytoperm Cytoperm Buffer 4 qA1 0 00000000 Perm Wash Buffer 25 ml 1 I 0 Perm Wash Buffed 0000m 110x0000 OO00Perm Wash 1
23. 554719 554720 559336 554498 559337 554706 559330 554707 554574 554575 559329 554576 557020 559325 554571 559328 554736 554506 554507 555042 554700 554701 559327 554702 554551 554552 559326 555049 554665 554666 559324 555033 555039 20794A 20795A 20795X 18555A 18555X 20705A 20705X 20709A 18994A 18995A 18995X 18999A 23275A 23275X 18965A 18965X 20985A 18594A 18595A 23125A 20664A 20665A 20665X 20669A 18904A 18905A 18905X 23175A 20524A 20525A 20525X 23025A 23105A Format Size FITC PE PE PE PE 0 1 mg 0 1 mg 100 Tests 0 1 mg 100 Tests 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg 100 Tests 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 0 1 mg 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg New Cat No Old Cat No 554729 554730 559323 554732 559322 559968 554512 554513 559321 554514 554556 New Cat No 559810 554427 554428 554429 554377 554378 554383 554388 554389 554435 554436 554395 554396 554401 554466 554467 554468 Specificity Clone Isotype Human Cytokines and Chemokines Continued MIP 10 11A3 Mouse lgG2a RANTES 205 Mouse IgG1 Thioredoxin 2G11 TRX Mouse IgG1 TNF a MAb 11 Mouse IgG1 359 81 11 Mouse IgG1 Specificity Clone Isotype Mouse Cytokines and C
24. FLA FL4 o FL3 OD faur Coar raram 95 2 FL3 OFL200 00000000 OOO PE antidL 10 0O0000 8000000000 0FL2 OFL30000 7 AAD 2 10 4 anti IL 10 2 2 9 1 trm 2 22 ee cee goes wees 5 Em mp ijt B Pi UMP 5 rn Dn 5 Pa W mr p oji P3 i m san we i laa x FL Baa 1 a i Dn 5 P ri i 8 Lin FL2 36 3 FLI FL2 4 4 FL3 91 24 2 FY rL14 w D 5 Lim FL3 1 97 FLA FL4 737 ifr BDM Perm 10 2 anti BrdU 7 anti TNF o 2 anti IL 100 0000000 9 anti TNF of 0000000 UT TNF oP OOO FL2Q0
25. 000000 OPE anti L 105 t 0 DO Xd DO BD ODCUGU UD UD U 1 2 1 9 2 7 1 FL2 PMT voltage OOOOUUFL200000 00000070900000 5 0 FL20 FL2 O FL30 00000000098 UOUOU0 19 820 B40 000000000 OOO 2 2 gi OOO Braun og EE HE EEE ASAE EREEREER E BERE ENAH
26. 20 21 22 23 24 25 26 27 Penit C 1986 in vivo thymocyte maturation BrdU labeling of cycling thymocytes and phenotypic analysis of their progeny support the single lineage model J Immunol 137 2115 2121 Thoman M L 1997 Early steps in T cell development are affected by aging Cell Immunol 178 117 123 Tough D F and J Sprent 1994 Turnover of naive and memory phenotype T cells J Exp Med 179 1127 1135 von Boehmer H and K Hafen 1993 The life span of naive alpha beta T cells in secondary lymphoid organs J Exp Med 177 891 896 Schittek B K Rajewsky and l Forster 1991 Dividing cells in bone marrow and spleen incorporate bromodeoxyuridine with high efficiency Eur J Immunol 21 235 238 Rocha B C Penit C Baron F Vasseur N Dautigny and A A Freitas 1990 Accumulation of bromodeoxyuridine labeled cells in central and peripheral lymphoid organs minimal estimates of production and turnover rates of mature lymphocytes Eur J Immunol 20 1697 1708 Westermann J S Ronneberg F J Fritz and R Pabst 1989 Proliferation of lymphocyte subsets in the adult rat a comparison of different lymphoid organs Eur J Immunol 19 1087 1093 Robey E D Chang A Itano D Cado H Alexander D Lans G Weinmaster and P Salmon 1996 An activated form of Notch influences the choice between CD4 and CD8 T cell lineages Cell 87 483 492 Carayon P and A Bord 1992 Identification o
27. 3 555111 555110 Positive for IL 2 IFN y Positive for IL 3 IL 4 IL 10 IL 13 GM CSF Positive for IL 1a IL 1B IL 6 IL 12 TNF a Positive for IL 8 GRO IP 10 MCP 1 Old Cat No 19701V 19731V 19991V 19741T 19801T 19761T 19771T Old Cat 192 EEP 19221T T923TT 19241V 19251 19281T 19291T 19341V 19321T 25011V 25071V 25051V 25041V No Format Standard Standard Standard Standard Standard Standard Standard Format Standard Standard Standard Standard Standard Standard Standard Standard Standard Standard Standard Standard Standard New Cat No 555061 555062 555063 555064 Size gt Jg gt Ug gt Jg 10 ug 10 ug 10 ug 10 ug Size 20ug 10 ug 10 ug 5 Ug 5 Ug 10 ug 10 ug 5 Ug 10 ug 5 Ug 5 Ug 5 Hg 5 Ug Old Cat No 232617 252024 232637 232644 29 Intracellular Cytokine Positive Control Cells Description Cytokines Expressed Mouse MiCK 1 Positive for IL 2 IFN y MiCK 2 Positive for IL 3 IL 4 IL 10 GM CSF TCA3 MiCK 3 Positive for IL 1a IL 6 IL 12 MCP 1 TNF a Rat RICK 2 Positive for IL 4 IL 10 GM CSF IFN y TNF a Specificity Clone New Cat No Old Cat No Isotype Controls Mouse IgG1 MOPC 21 554679 20604A 554680 20605A 559320 20605X 554681 20609A Mouse lgG2a G155 178 554647 20074A 554648 20075A 559319 20075X Mouse IgG2b 27 35 555057 23244A 555058 23245A Rat IgG1 R3 34 554684 20614A 554685 20
28. 615A 559318 20615X 554686 20619A Rat IgG2a R35 95 554688 20624A 554689 20625A 559317 20625X 554690 20629A Rat IgG2b A95 1 556923 20184A 556925 20185A 556924 20189A Hamster IgG 4235 2356 554710 20724A 554711 20725A 30 New Cat No Old Cat No 554652 554653 554654 555094 Format 20131Z 2013527 201932 241427 Size 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 0 1 mg 100 Tests mg 0 1 mg 100 Tests mg mg mg mg mg mg oolooo o 1 1 1 1 1 1 Intracellular Cytokine Flow Cytometry Kits and Reagents Description Kits Cytofix Cytoperm Kit New Cat No Old Cat No 554714 554715 2075KK 2076KK Cytofix Cytoperm Plus with Golgi Stop Cytofix Cytoperm Plus with GolgiPlug 555028 2040KK 2300KK Human Intracellular Cytokine Staining Starter Kit 559302 2041KK Mouse Intracellular Cytokine Staining Starter Kit 559311 BrdU Flow Kit FITC Description Reagents Buffers Cytofix Cytoperm Buffer New Cat No 554722 554723 559619 Old Cat No 2090KZ 2354KK Size 125 mLs Perm Wash Buffer 10X 554724 2091KZ 2092KZ 100 mLs 0 7 mL GolgiStop containing monensin 555029 2301KZ 1 0 mL GolgiPlug containing brefeldin A Cytofix Buffer 554655 2014KZ 100 mLs Pharmingen Stain Buffer FBS 554656 20151E 500 mLs Pharmingen Stain Buffer BSA 554657 20161E 500 mLs 31 Secti
29. AX 03 6206 7934 http www fujisawa co jp reagent 03 5805 9960 64 035 01 RO 0111 000 5 540
30. f DNA replicating lymphocyte subsets using a new method to label the bromo deoxyuridine incorporated into the DNA J Immunol Meth 147 225 230 Gonchoroff N J J A Katzmann R M Currie E L Evans D W Houck B C Kline P R Greipp and M R Loken 1986 S phase detection with an antibody to bromodeoxyuridine Role of DNase pretreatment J Immunol Meth 93 97 101 Shapiro H M Practical Flow Cytometry 3rd Edition Wiley Liss New York Gray J W and Z Darzynkiewicz Eds Techniques in Cell Cycle Analysis Humana Press Clifton New Jersey 33 34 Notes Notes 35 amp BD uuu 8 5 26 6 107 0052 u 1 uuu Es BD Pharmingen milli ll 101 0043 10 2 T fry 0120 8555 90 TEU 03 5256 5311 FA 03 5256 5370 Fax 024 593 5761 541 8514 3 4 7 Life Science Support 3150120 7752 77 TEL 06 6206 7890 F
31. hemokines IL 10 ALF 161 Hamster IgG IL 2 JES6 5H4 Rat IgG2b IL 2 S4B6 Rat IgG2a IL 3 MP2 8F8 Rat IgG1 IL 4 BVD4 1D11 Rat IgG2b IL 4 11B11 Rat IgG1 IL 5 TRFK5 Rat IgG1 IL 6 MP5 20F3 Rat IgG1 IL 10 JES5 16E3 Rat IgG2b IL 12 p40 p70 C15 6 Rat IgG1 554479 554480 20954A 20955A 20955X 20975A 20975X 23714A 18644A 18645A 18645X 18649A 18915A Old Cat 23405A 18174A 18175A 18179A 18004A 18005A 18015A 18034A 18035A 18195A 18199A 18055A 18059A 18075A 18434A 18435A 18439A 18495A 18499A No Format FITC PE PE Format PE FITC APC FITC PE FITC PE PE APC PE APC PE FITC APC PE APC Size 0 1 mg 0 1 mg 100 Tests 0 1 mg 100 Tests 0 1 mg 0 1 mg 0 1 mg 100 Tests 0 1 mg 0 1 mg Size co e OO mx Eee eee axe I I I I j i 27 New Cat No Old Cat No 559502 554405 554406 554411 554412 554413 554443 554418 554419 554420 559503 New Cat No 555088 559498 559499 555092 554443 Specificity Isotype Mouse Cytokines and Chemokines Continued IL 17 Rat IgG1 GM CSF Rat IgG2a IFN y Rat IgG1 MCP 1 Hamster IgG Rat IgG1 Hamster IgG Specificity Isotype Rat Cytokines and Chemokines IL 4 Mouse IgG1 IL 10 Mouse IgG2b IFN y Mouse IgG1 GM CSF Mouse IgG1 MCP 1 Hamster IgG Hamster IgG Pig Cytokines and Chem
32. okines IFN y Mouse IgG1 559503 559812 Recombinant proteins useful as specificity controls for intracellular cytokine flow cytometry Specificity Human IL 10 IL 2 IL 3 IL 4 IL 5 IL 6 IL 8 28 New Cat No 554601 554603 554604 554605 554606 554607 554609 Old Cat No 19601V 19621T 19631T 19641V 19651V 19661T 19681P 23295A 18094A 18095A 18114A 18115A 18119A 18245A 18134A 18135A 18139A 23355A Old Cat 24055A 24095A 20444X 20445X 24135A 18245A 23355A 20455A Format Standard Standard Standard Standard Standard Standard Standard No Format Size PE FITC PE FITC PE APC PE FITC PE APC PE Format PE PE FIG PE PE PE PE PE Size gt Ug 10 ug 10 ug gt Ug gt Jg 10 ug 20 ug GOS eee Size 0 1 0 1 100 Tests 100 Tests 0 1 0 1 0 1 0 1 mg mg mg mg mg mg Specificity Human Continued IL 10 IL 13 IL 16 GM CSF MIP 10 Specificity Mouse IL 2 L 3 IL 4 IL 5 IL 6 IL 10 GM CSF MCP 1 TNF a Rat IL 4 IL 10 GM CSF MCP 1 Intracellular Cytokine Positive Control Cells Cytokines Expressed Description Human HiCK 1 HiCK 2 HiCK 3 HICK 4 MCP 3 MIG MIP 1a New Cat No 554611 554614 554637 550068 554622 554618 554619 New Cat No 550069 554579 550067 554581 554582 550070 554586 554590 554589 555107 55511
33. on 8 ELE ES Sasaki K T Murakami and M Takahashi 1989 Flow cytometric analysis of cell proliferation kinetics using the anti BrdUrd antibody Gan To Kagaku Ryoho 16 2338 2344 Miltenburger H G G Sachse and M Schliermann 1987 S phase cell detection with a monoclonal antibody Dev Biol Stand 66 91 99 Vanderlaan M and C B Thomas 1985 Characterization of monoclonal antibodies to bromodeoxyuridine Cytometry 6 501 505 Gratzner H G and R C Leif 1981 An immunofluorescence method for monitoring DNA synthesis by flow cytometry Cytometry 1 385 393 Lacombe F F Belloc P Bernard M R Boisseau 1988 Evaluation of four methods of DNA distribution data analysis based on bromodeoxyuridine DNA bivariate data Cytometry 9 245 253 ean P N F Dolbeare H Gratzner G C Rice and J W Gray 1984 Cell cycle analysis using a monoclonal antibody to BrdUrd Ce Tissue Kinet 17 427 436 Toba K E F Winton and R A Bray 1992 Improved staining method for the simultaneous flow cytofluorometric analysis of DNA content S phase fraction and surface phenotype using single laser instrumentation Cytometry 13 60 67 Sasaki K S Adachi T Yamamoto T Murakami K Tanaka and M Takahashi 1988 Effects of denaturation with HCl on the immunological staining of bromodeoxyuridine incorporated into DNA Cytometry 9 93 96 Lakhanpal S N J Gonchoroff J A Katzmann and B S Handwerger 1987 A flow cytofluorome
34. tric double staining technique for simultaneous determination of human mononuclear cell surface phenotype and cell cycle phase J Immunol Meth 96 35 40 Houck D W and M R Loken 1985 Simultaneous analysis of cell surface antigens bromodeoxyuridine incorporation and DNA content Cytometry 6 531 538 Moran R Z Darzynkiewicz L Staiano Coico and M R Melamed 1985 Detection of 5 bromodeoxyuridine BrdUrd incorporation by monoclonal antibodies role of the DNA denaturation step J Histochem Cytochem 33 821 827 Holm M M Thomsen M H yer and P Hokland 1998 Optimization of a flow cytometric method for the simultaneous measurement of cell surface antigen DNA content and in vitro BrdUrd incorporation into normal and malignant hematopoietic cells Cytometry 32 28 36 Mehta B A and V C Maino 1997 Simultaneous detection of DNA synthesis and cytokine production in staphylococcal enterotoxin B activated CD4 T lymphocytes by flow cytometry J Immunol Meth 208 49 59 Endl E P Steinbach R Kn hel and F Hofst ter 1997 Analysis of cell cycle related Ki 67 and p120 expression by flow cytometric BrdUrd Hoechst 7AAD and immunolabeling technique Cytometry 29 233 241 Dolbeare F H Gratzner M G Pallavicini and J W Gray 1983 Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine Proc Natl Acad Sci USA 80 5573 5577 32 16 17 18 19
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