免疫機能分析マニュアル 肉用鶏特別飼育分科会
Contents
1. Anti microbial growth promoter2. NK 3 T B T B 1 2 3
3. lt 3 1 i BARE Ele os A A i i fa 2 3 4 Th1 1848 NEE SS J BSS SSRIS iman gt Thommen sated eee nee CD8T Th2 aie 3 E 3 HELL BELU Kh 3 5 3
4. 3 4 Matsushita K Takahashi K Effect of Adequate or Marginal Excess of Dietary Methionine Hydroxy Analogue Free Acid on Growth Performance Edible Meat Yealds and Inflammatory Response in Female Broiler chickens J Poult Sci 44 2007 10 SRBC BM T Sheep Red Blood Cell T 1 a 5ml 4500 ere meres EE wako 5g 6000 BSA 96
5. 8 2 000 2 000 8 250 250 No 2 Oo a ee as 10 11 12 13 14 2 4 8 1024 2048 4096 8192 9B OR YOR awww x a x F TA S N i 25 25 25 25 25 25 25 25 25 8 25 25 25 rrr 25 25 25 25 50 50 50 rr 50 50 50 50 50 1024 45 end tite DE 2 8
6. 1 2EK 5 m ie ei al belie RR ium PE A U JE 50ml 5000 ND 2 gt EE eat gt D 3 2 500 rpm 10 packed cell 0 59 D HA 500 09ml 09 08 IN IN 0 1ml 0 1 0 2 No 1 2 3 4 5 6 1000 2000 4000 8000 16000 50u 50 50 50 50 50 RSS ee a Sa eee 500 50u 50 50 50 50 50u 50 50 50 50 50 ES HA
7. 4 5 6 4 4 3 Ata T
8. DMEM imi 10cell ml 100 DMEM 1980u 20u 5 x 10 cell ml DMEM x 10 cell ml Y O 10mg ml DMEM 600u 1 2 D 200u 1 1 x eh TT Ir Beormomeemvicts 94 YYIM XI h 0 cell 5 CO 37C D
9. II ou oe 19 AAR AB He DP PUL OABOFSABCOBSIL 1 ANSEL To 2 470 mie a a a ae 15 Pisin east H21 5 2 3 4 20 EEEn 1 gt gt gt gt gt gt St eee ee eee A ee 21 1 DIMEM 500ml 1 500
10. 3 4 18 eee oe eee ee eee ee eee gt ee 2 gt L a c d B y _ s s gt a gt PE SOE HOE Oe Oe Oe Oe OO Oe OH OO ew Om OORA RBS A1 H LTS cn
11. _ Cost nmol Lys kg ad Fyne gaia 46 91 i 5 we 66 70 0 386 111 547 ais sits i Lys 1 17 6 71 5 E Rosales From Humphrey and Klasing 2004
12. 30 30 1000 3 7 1 Saat BZ ER _ 1 1030 500g 1 1310 500g 1 2760 100g O 0 1M pH5 4 1 5M 27mM A 0 5g 0 1M 100ml 3 0 5g 100mll 27mM 27mM
13. 1 Mraka seet aa ma ee BETHYL ELISA Quantitation BETHYL 1Kit 38 000 ELISA Kit HRP Chicken E30 103 ON 1000 ELISA Starter Accessory BETHYL 1Kit 42 000 Package E101 450nm 20 C 2 1 ELISA Starter Accessory Package BETHYL E101 A1 Coating Buffer A2 Wash Solution A3 Postcoat Solution A4 Sample Conjugate Diluent A5 Enzyme Substrate A6 2N H2SO4 2 Capture Antibody 3 4 5 HRP Detection Antibody 6 Enzyme Substrate Reaction 7 450nm 3 IgA 4 BETHYL ELISA Quantitation Kit Chicken E30 103 BETHYL ELISA Starter Accessory Package E101 29 TBARS
14. D 80C 5 29 6SRBC 1096BA 2 1 2 4m HE D 2 0 0596BSA PBS 1 2 4ml HE 25u 1 25u 12 25u 6 ERIZ 296SRBC 109 BA 37C1 BA 1 ARE D 5096 2 12 ND HD COMER MR AES HR
15. No 1 2 3 4 5 6 7 8 1 2 4 8 16 32 64 25 25 25 25 25 25 25 RS DU SS 2 NR SN Se 2 Re 500 25u 1 25 25 25 25 25 25 50 50 50 50 50 50 50 50 1 8 H17 14 1 vy 10mlx 6 g aa 9 520 100 BEI m 390 1ml 100 1 on 280 100 ml 29Gx 6 000 1 2 500g 640 50ml Centrifuge Tube IE 6 300 23 560 H21 5 OE CR
16. 1 x 2 1 2
17. O T O OHL T T x NK A O NO IgA IgG OBER SUE JUANKORE OSS SRBC BM ND MFHT LOA BMH REO Ale 1 mM Hepes Eagles s MEM T 4000 L glutamine ako 5 ako ako ako IE GMA ako 500ml HER I TERUMO100 19G xi HM TERUMO 1004 8700 lei ml NIPRO 1 ALOKA 41 LUMINESCENCE REA
18. H X 0 URR 1 SRBC BM ND H L F Mn IgA TBARS LSP NO p ISU MIS She aR O RELI Alle BAO lo A
19. PBS PBS 3 500u 10 D 500u 15 1 2 1000 5u
20. 2 0 1M 2ml 2 blank and reaction Doim 37 5 A 1m 5 50u 30 50ul 530nm 3 g 0 752 x A Ag 4 Matsushita K Takahashi K Effect of Adequate or Marginal Excess of Dietary Methionine Hydroxy Analogue Free Acid on Growth Performance Edible Meat Yields and Inflammatory Response in Female Broiler chickens J Poult Sci 44 2007 Matsushita K Takahashi K Effect of Dietary Protein Source and Antibacterial Agent on Growth Performance Edible Meat Yield and Certain Stress Response in Female Broiler Chickens J Poult Sci 41 2004 Shimosato F Studies of analytical methods for ceruloplasmin in Serum 1 A comparison of the ferrous oxidase reaction and the diamine oxidase reaction in quan
21. 7 10mg m1 V PIV IVT I bic Vv 5 X 106well ml wm Vv v CO 37 C WRES V w F Vv V NY CO 37C 1 V V ke iR BE V Vv SAS SYA SR T lt RARAKYIR OR gt BHI 50ml co k 12 1ml D 37C 24 50ml 1ml 80C SEI EAFORE gt BHI 50ml E coli k 12 1ml D 37C 24 HH 0 59 BHI 50ml 0 25mD 50m 2 400
22. 8 550nm well 9 222 Nitrite Nitrite 3 in vitro 4 4 47 50 2004 NO 30
23. 1 BAK THRE 500ml 1 8800 ANEIROS 1 4600 1 1 3 3 mA 1 2 300 25ml 500ml Na 25g AIA oo 500m1 500m pl 1 7 000 16 5X 130mm on TE ae 6 GeO BERR VortexMixer 0 6796TBA 5096 1 1 100ml 0 67g 2 STD 5nmol ml 55mg 1 1 3 3 50ml 50ml 0 1ml 100ml 100ml 10 10g Na
24. BESTE 1 ml y 400u g 10 1 Im 4 0 25ml 15 D 2 m y 400u g 0 1ml 0 1ml 3 O 24 48 D
25. 5 8 200u 1 300u 15 27 1000 3 3 x100 gt 8 28 IgA
26. 100ml ETS N 12H SO 36N M 12 n 30 2 1 STD 100u H 0 3 9ml TBA 1 0ml FMA FRE STD 5nmol ml 0 250 0 125 0 0625 3 1 10 0 5ml N 12H SO 4 0ml 20u 2 5min 3500rpm10min 3 N 12H SO 2 0ml 10 0 3m MAH KRH 4 5min 3500rpm10min 5 H 0 4 0ml TBA 1 0ml 6 60min 8n 5 0m 2min 3000rpm10min 9 STD 515nm 530 570nm 553nm 550nm
27. 4 16 sa A Ss 1 a H L EE 1 MICRO SLIDE GLASS 100 ow eee A a x if L 3y g 8080 AA 100 25ml 23GX 1 mi 1 250 IMMERSION OIL FOR 6 H22 5 D 1 1 5 AF Ae 1 1 5 D NA 1 1 5 GST T
28. 15m 1000rpm 7min at 4 C 5ml RPMI 1640medium 1000rpm 7min at 4 C 5ml RPMI 1640medium 1000rpm 7min at 4 C 2 2ml 733u X 3 1 Chicken serum 10 FBS 1 6 RPMI 1640medium cell count Burker Turk Type 0 2ml 2 0ml 0 3 PBS 1 1 Count LAL Cell Count 2 0 x 107cells ml Chick PBMC 2 0 x 10 cells ml 200p well 48well plate Well 200
29. 50 LY Fa 16500 V J 2 5mD FILE 100 AY 2400 1ml 2 gt 1 1096 0 05ml 2 D 2000 2500rpm 20 D O D 3 D 1 x ml 9ml x 9 2 296BA D BA 2000 2500rpm 20 D 200u 49 200u 9 8ml 11 3 1 2 gt 4 21 28 0 1m 4 5
30. DMEM 500ml x 20 Latex beads polystyrene 1 1u m SIGMA 10 600 ee 15m 500 28 500 I 8 16 12 960 64 2 rasen 2589 om som Feaee ESRI lisoo FT PALE PC BETO Fraea moa om fio ee ee si AR wok o wR fiom 0 08x1 em pog 49 Escherichia colik 12 NBRC 4200 wase OoOO o aE I E 0 PBS 1 EE 6 0 REAR 5 CO EE DE EE EE PBS Phosphate Buffered Saline NaCl 8 0g KCI 0 2g Na HPO 1 15g KH PO 0 2g 1L 5ml DMEM DMEM 5ml 22 2 WA h NY
31. 3 TBARS 4 1983 2000 2005 l LPS 1 Lipopolysaccharides fro m Escherichia coli O12 7 B8 L3129 100mg 2 Lipopolysaccharides from Escherichia coli 0127 B8 ikiq 32 Ria 7K IIIB AeL 500u g ml 500u g m keg 3 LPS 4 K Takahashi T Mashiko and Y Akiba Effect of Dietary Concentration of Xylitol on Growth in Male Broi ler Chicks During Immunological Stress Poultry Science 9 743 747 2000 K Takahasi Y Akiba and K Tamura Effect of Repeated Sephadex Injection with or without Escherichia coli Lipopoiysaccharide on Growth Immunocompetent Organ Weight and Plasma Alpha 1 Acid Glycoprotein Concentration in Broiler Chicks Reared in Different Stocking Density
32. 9 5u 5u X100 gt 5 5g 5u 26 lt BR APT YIRAE XE gt O II 200u n 2 D 5 CO2 37 C DMEM A 3 0 039 DMEM 12ml 40u 1 200u 5 CO 37 C 1 PBS
33. 0rpm 10 10 C 4000rpm 10 10 C 2 80 C D AD 10mg ml DMEM 37 C CO2 lt UL RORE gt D 4ml EDTA 15 30 C D 3 5ml 1800rpm 20 15 15 30 C D PNGV 2 RS 6 DMEM 1ml 15m FEL BECHER TS 2 1400rpm 10 15 C 24 D
34. Anim Sci J 71 3 268 273 2000 32 NO 1 i 7 oy 100 21 000 ete Ficool paque Amersham Biosiences 100ml x 6 33 000 RPMI 1640 SIGMA 500ml 6800 El DYIARYZ A 200u 96 5 7 8 800 I x 10 HA Long tip 96 _ WATSON UN ha me lm waes mm sa mm tem em sml HK N A YE TIN SNS 550nm 5 CO2 33 2 Chick PBMC peripheral blood mononuclear cell 7ml RPMI 1640medium 15m 2 5ml Ficool paque 15m 2000rpm 10min at 20 C PBMC 5ml RPMI 1640medium
35. DER 2 2 1 5mM Hepes Eagle s MEM E MEM Eagle sMEM9 4g L glutamine0 292g DW1000ml l 10 NaHCO G pH7 4 NaHCO 20ml pH7 4 0 45u m 20 C Luminol 50mg 25u DW25u 1 50C 15min x 3 m Scie ZymosanA 100mg 0 15MNaCl 10ml nice 10min aii 2 2000rpm 10min re Im ae 37 C 30min wo 10 PBS 2 2000rpm 10 5mM Hepes Eagle s MEM10ml 100mg WL 20C 2 50u E MEM 450u 10u 10u
36. titative determination of serum ceruloplasmin Shinshu unv 1 984 Sunderman FW Jr Nomoto S Measurement of human serum ceruloplasmin by its p phenylenediamine oxidase activity Clinical Chemical 16 1970 a 1 1 a 1AG 1 47 250 100 2 B 5u 37C 48 5u 37 C 48
37. u 5 0u L 5 0u L 20h 5 CO2 35u L NO assay 34 Nitrite 4C 200u M Griess 4C Griess1 7 5 phosphoric acid 1 0 sulfanil amid DW Griess2 0 3 DW 1 2 1 2 96well plate 550nm 1 35u L 96well plate 2 Nitrite O u M 5 u M 10 u M 20 u M 35 u M 70 u M ook 0 2 0 5 0 10 0 17 5 35u L Nitrite DW 100 97 5 95 0 90 0 82 5 65u L 3 Nitrite 35u L assay well 4 Griess Griess1 Griess2 1 2 5 Nitrite well Griess 35u L 6 10 D
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