Download Ultrospec 3100 pro Spectrophotometer
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0.76×(Abs λ260 ) ç¹æ®ãªã¿ã³ãã¯è³ªã«åŒã察å¿ãããããã«ã¯ãæ¿åºŠæ¢ç¥ã¿ã³ãã¯è³ªã 260 nm ããã³ 280 nm ã§åžå 床枬å®ããçžé¢ä¿æ°ãéç®ããããšãã§ããŸãããã®å ŽåãX ãš YïŒç® çã®çžé¢ä¿æ°ïŒã倿°ãšããŠè§£ãé£ç«æ¹çšåŒã«ãªããŸããXå€ãè² ã®ä¿æ°ã«ãªãå Žå ã¯ãX å€ããŒãã«èšå®ããŠãã ããïŒ260 nm ã§ã®åžå 床ã¯ãã¿ã³ãã¯è³ªæ¿åºŠã«å šã å¯äžããªãããè² ã®ä¿æ°ãçããå ŽåããããŸãïŒ ããŸãå¿ èŠã«å¿ããŠããã¯ã°ã©ãŠ ã³ãåžå 床ãè£æ£ããããšãã§ããŸãã * åèæç® : Warburg and Christian, Biochemisches Zeitung 310, 384 (1941). ã¿ã³ãã¯è³ªæ¿åºŠ =ãã¡ã¯ã¿ãŒ1 ×(Abs λ280 - Absλ320 ) - ãã¡ã¯ã¿ãŒ2 × (Abs λ260 - Abs λ320 ) λ 320ïŒããã¯ã°ã©ãŠã³ãïŒã¯ãªãã·ã§ã³ãªã®ã§ã䜿çšãããã©ããéžæã§ããŸãã ãã¡ã¯ã¿ãŒã®èšå®å€ã¯å€æŽå¯èœã§ãã çŽæ¥Î» 280 ã§ã¿ã³ãã¯è³ªã枬å®ããå Žåããã¡ã¯ã¿ãŒ 2 = 0.00 ã«èšå®ãã¹ã¿ã³ããŒã ãšã㊠BSA(bovine serum albumin)ã䜿çšããå Žåããã¡ã¯ã¿ãŒ 1 = 1.115 ã«èšå®ãã ããšã§ 0 ⌠0.8mg/ml éã¯çŽç·çãªçµæãåŸãããŸãã Background on âš Factor 1 ãã¡ã¯ã¿ãŒ 1 ãèšå®ããŠãã ããã ïŒåæèšå®å€ 1.55 mg/mlïŒ Factor 2 ãã¡ã¯ã¿ãŒ 2 ãèšå®ããŠãã ããã ïŒåæèšå®å€ 0.76 mg/mlïŒ yes, Save Method Load âš âš âš yes, âš âš ããã¯ã°ã©ãŠã³ãè£æ£ã¯å¿ èŠã§ããã ãã®ã¡ãœãããä¿åãããšã㯠enter and Result Sample run ãªãã¡ã¬ã³ã¹èšå® run ãµã³ãã«æž¬å® ïŒ ãªãã¡ã¬ã³ã¹ã®æ¬¡ã®ãµã³ãã«ããµã³ãã«çªå·1ãšãªããŸãã è¡šç€ºã®æ°åã¯ãµã³ãã«ã§ãããã»ã«ã®äœçœ®ã§ã¯ãããŸãããïŒ 21 Empirical 280, 205 nm å¿ èŠã§ããã°ä¿åã¡ãœãããšããŠä¿åã§ããŸãã enter ã¿ã³ãã¯è³ªæ¿åºŠïŒmg/mlïŒã¯æ¬¡ã®å®éšåŒã«ããæšå®ã§ããŸãã ã¿ã³ãã¯è³ªæ¿åºŠïŒmg/mlïŒïŒ27 + (120×A280/A205) âš Scopes, R. K (1974) Measurement of protein by spectrometry at 205 nm Anal. Biochem. 59, 277-282 Save Method Load and If yes, âš åèæç®ïŒ run ããŒãæŒããŠãªãã¡ã¬ã³ã¹ããšãã run ããŒãæŒããŠãµã³ãã«ã枬å®ããŸãã enter Concentration Ratio ãš ã衚瀺ãããŸãã Empirical 215, 255 nm å¿ èŠã§ããã°ä¿åã¡ãœãããšããŠä¿åã§ããŸãã enter ã¿ã³ãã¯è³ªæ¿åºŠïŒµg/mlïŒã¯æ¬¡ã®å®éšåŒã«ããæšå®ã§ããŸãã ã¿ã³ãã¯è³ªæ¿åºŠïŒmg/mlïŒïŒ144×A215/A225) âš Save Method Load and If yes, âš åèæç®ïŒ Waddel, W.J (1956) A Simple UV Spectrophotometric method for the determination of protein. J. Lab. Clin. Med. 48, 311-314 run ããŒãæŒããŠãªãã¡ã¬ã³ã¹ããšãã run ããŒãæŒããŠãµã³ãã«ã枬å®ããŸãã enter Result ã衚瀺ãããŸãã Info æ°å€ãã®äžè¬çãªã¿ã³ãã¯è³ªã«ã€ããŠãA280 ã®æž¬å®å€ããæ¿åºŠïŒmg/mlïŒãžæç®ããåŒããã ã³ã¢ããé žã«ã€ããŠã®äžè¬çãªæ å ±ãåç §ã§ããŸãã 22 Applications enter Wavescan enter æ³¢é·ã«å¯Ÿããåžå 床å€åã®ã°ã©ãã¯ãåžå ã¹ãã¯ãã«ãšããŠç¥ãããŠããŸããã ç©è³ªã®ç©ççç¹åŸŽã®1 ã€ã§ããã宿§åæãå®éåæã®ææ®µãšããŠæçšã§ããåžå ã¹ãã¯ãã«ã¯ãååäžã§èµ·ããããçš®ã ã®é»åé·ç§»ã«ããçãããã®ããŒã¯ã¯ ïŒæº¶æ¶²äžã§ã¯ïŒå¹ åºããªããŸãã ã¹ãã¯ãã«ã®å°é¢æ°ããã¯æ¬¡ã«æããæ å ±ãããã«åŸãããŸãã 1次埮åã§ã¯ãå¯ æ¥ããå€ãã®ããŒã¯ã®åå®ãã§ããŸãã 2 次埮åããã¯ãããŒã¯ã·ã§ã«ããŒïŒå± æ²ïŒã®åå®ãå¯èœã§ãããããŠ4次埮åã¯ãåæã«å€ããŒã¯ãšå±æ²ãäž¡æ¹ã®åå® ãã§ããŸãã Set up âš âš Start λ éå§æ³¢é·ãå ¥åããŸãã End λ çµäºæ³¢é·ãå ¥åããŸãã Scan Speed* âš Data Interval* âš Reference scan If yes, âš Save Method If yes, Load âš âš âš âš âš ã¹ãã£ã³é床ãéžæããŸã ïŒFast, Medium, Slow)ã ããŒã¿ã枬å®ããééãïŒ1.0ã0.5 nmïŒ éžæããŸãã ãªãã¡ã¬ã³ã¹ã®ã¹ãã£ã³ã¯èšæããŒã¹ã© ã€ã³ã«ãªããŸãã enter run ïŒãªãã¡ã¬ã³ã¹èšå®ããå ŽåïŒ, and * ãããã®ã¹ãã£ã³é床ïŒnm/minïŒãšæž¬å®æ³¢é·éé æž¬å®æ³¢é·éé Fast é«é 1.0nm 0.5nm 1800 900 Medium æ®é 750 375 Slow 250 125 äœé 23 Graph 以äžã®æäœã§ãã€ã³ããå ¥åããã°ãæ£ç¢ºãªäœçœ®ã§ãºãŒã ãè¡ããŸãã éå§æ³¢é· âš çµäºæ³¢é· âš Y 軞ã®ãªãŒãã¹ã±ãŒã«ãè¡ãå Žåã«éžæã㟠ãã æå€§åžå 床ãéžæããŸãã æå°åžå 床ãéžæããŸãã âš âš âš âš âš âš enter ãºãŒã ããã«ã¯ ãºãŒã ããã«ã¯:: ããŒã§ãºãŒã ãããããŒã¯ã«ã«ãŒãœã«ãç§»åããŸãã ãæŒããšæ»ããŸãã ãé£ç¶ããŠæŒããŠãã ããã âš Overlay âš å°é¢æ°ãéžæããŠã¹ã±ãŒãªã³ã°æ©èœã䜿ã£ãŠã¹ãã¯ãã«ã«éãã ããšãã§ããŸããã¹ãã¯ãã«ããŒã¿ã¯ã¹ã ãŒãžã³ã°ã% T ãžã®å€æ ãã§ããŸãã âš Type Offã1stã»2ndã»4th DerivativeãSmoothã Enhanceã%TransmissionããéžæããŸãã âš Scale âš Offset å®éšããã³ããªã³ãã¢ãŠãã®ããã«ïŒ¹è»žå€ ãå ¥åããŠL C D äžã§ãªãŒããŒã¬ã€ãè³é©å ããŸããScaleããã³Offsetã¯Typeã®ã¿ã«åœ±é¿ ããŸããå ¥åããæ°å€ãšT y p e ã®åãã¹ã±ãŒ ã«ãšãªããŸããå¿ èŠã«å¿ããŠæšªåº§æšãã TypeãOffsetã«ããç§»åããŸãã } åã ã®ã¹ãã£ã³ã¯ãªãŒããŒã¬ã€ã§ããŸããã Peak Table âš âš â ãã®æ©èœã§ã¯ãããŒã¯ããŒãã«ãâã«èšå®ãããŠããæã«å šã¹ãã¯ãã«ã« ãããããŒã¯åžå åºŠãšæ³¢é·ã®å€ããªã¹ãã¢ããããŸãããããŒã¯å¹ ã 1 5 n m 以äžãããã®ã«ã€ããŠã¯ãããŒã¯ãšããŠèªèããŸãããèªèãããªãããŒã¯ 㯠ããŒã䜿ã£ãŠã«ãŒãœã«ãç§»åããŠç°¡åã«åå®ã§ããŸããå¹³ã ãŸãã¯å¹ åºã®ããŒã¯äžã®æ³¢é·å€ã¯æ£ç¢ºã§ã¯ãªãã®ã§ãïŒâïŒã§ç€ºãããŸãã çµæã¯å€æŽã§ããŸããã 24 Applications enter Multiwave enter Abs Ratio enter Absorbance RatioïŒåžå 床æ¯ïŒã¯ååçç©åŠåéã§å©çšãããŠããæ žé žã®å®éãçŽåºŠ 詊éšã«çšããŠãã ããã ⚠λ2 æ³¢é·2ãå ¥å Background on If yes, âš æ³¢é·1ãå ¥å ããã¯ã°ã©ãŠã³ãè£æ£ãå¿ èŠãªãã° If yes, λB ããã¯ã°ã©ãŠã³ãã®æ³¢é·ãå ¥å Factor æ³¢é·1ã«å¯Ÿå¿ãããã¡ã¯ã¿ãŒãå ¥å ⚠⚠⚠⚠λ1 Units Save Method Load âš âš âš âš âš âš If yes, â åäœãéžæ åäœãéžæ enter and Ratio Concentration Sample run ãªãã¡ã¬ã³ã¹èšå® run ãµã³ãã«æž¬å® ïŒ ãªãã¡ã¬ã³ã¹ã®æ¬¡ã®ãµã³ãã«ããµã³ãã«çªå·1ãšãªããŸãã è¡šç€ºã®æ°åã¯ãµã³ãã«ã§ãããã»ã«ã®äœçœ®ã§ã¯ãããŸãããïŒ 25 Abs Diff enter Absorbance DifferenceïŒåžå 床差ïŒã¯åºè³ªã«å¹²æžããããã¯ã°ã©ãŠã³ãåžå 床ãè£ æ£ããå Žåã«çšããŠãã ããã λ2 æ³¢é·2ãå ¥å If yes, Background on âš æ³¢é·1ãå ¥å ããã¯ã°ã©ãŠã³ãã®æ³¢é·ãå ¥å Factor1 æ³¢é·1ã«å¯Ÿå¿ãããã¡ã¯ã¿ãŒãå ¥å Factor 2 æ³¢é·2ã«å¯Ÿå¿ãããã¡ã¯ã¿ãŒãå ¥å Units If yes, Save Method Load åäœãéžæ åäœãéžæ enter and Sample Result run â ããã¯ã°ã©ãŠã³ãè£æ£ãå¿ èŠãªãã° If yes, λΠ⚠⚠⚠⚠⚠⚠⚠λ1 âš âš âš âš âš âš ãµã³ãã«æž¬å® ïŒ ãªãã¡ã¬ã³ã¹ã®æ¬¡ã®ãµã³ãã«ããµã³ãã«çªå·1ãšãªããŸãã è¡šç€ºã®æ°åã¯ãµã³ãã«ã§ãããã»ã«ã®äœçœ®ã§ã¯ãããŸãããïŒ enter 3 Point net 3 point net 㯠amnioticæ¶²äžã®bilirubin 枬å®ãªã©ã®åŸããããŒã¹ã©ã€ã³ãç€ºãæ¿ç¶ãµã³ ãã«ã«ãããæ£ããããŒã¯é«ã枬å®ããå Žåã«çšããŠãã ããã λ1 ? λ2 ? æ³¢é·1 æ³¢é·2 æ³¢é·3 Factor ããŒã¯é«ã«å¯Ÿãããã¡ã¯ã¿ãŒ Units? Save Method Load If yes, åäœãéžæ åäœãéžæ ã¡ãœãããä¿åãããšã㯠â enter and Height run ⚠⚠⚠λ3 ? âš âš âš âš âš âš âš Area Sample ãµã³ãã«æž¬å® ïŒ ãªãã¡ã¬ã³ã¹ã®æ¬¡ã®ãµã³ãã«ããµã³ãã«çªå·1ãšãªããŸãã è¡šç€ºã®æ°åã¯ãµã³ãã«ã§ãããã»ã«ã®äœçœ®ã§ã¯ãããŸãããïŒ 26 enter Multiwave Multiwaveã§ã¯æå€§9æ³¢é·ã®åžå 床枬å®ãåæã«è¡ãããšãã§ããŸãããããã®æž¬å®å€ãšæå€§9 ã€ã®ç°ãªãä¿æ°ãçšããŠã2ã€ãŸã§ã®èšç®åŒã«åœãŠã¯ãããµã³ãã«ã®çŽåºŠãæ¿åºŠãæ±ããããš ãã§ããŸãã èšç®åŒã¯ãããããçŽãªã©ã«æžããŠçšæããŠãããšå ¥åã®éã«äŸ¿å©ã§ãã 2ããŒãžåŸã«å ¥åäŸãèšèŒãããŠããŸãã®ã§ãåç §ãã ããã enter Set up âš No. of λâs æž¬å®æ³¢é·ã®æ°ïŒæå€§9æ³¢é·ïŒ ⚠λ1 æž¬å®æ³¢é·1 âŠâŠ âŠâŠ Î»9 âš Integration Time æž¬å®æ³¢é·9 âš âš âš åæ³¢é·ãçãé ã«å ¥åããŠãã ããã ïŒNo. of λâs ã§å ¥åããæ°ã ã衚瀺ãããŸããïŒ 0.1ã1ã2ã5ç§ã®ãããããéžæãéåžž ã«èãïŒãŸãã¯æ¿ãïŒãµã³ãã«ã®å Žå㯠é·ãã«èšå®ããŠãã ããã enter Factors èšç®åŒã«çšããããã®ãã¡ã¯ã¿ãŒãä¿æ°ãå ¥åããŸãã ãµã³ãã«ã®åžéç K1 ä¿æ°1 âŠâŠ âš Dilition Factor, C âŠâŠ âš âš K9 ä¿æ°9 ãã€ãã¹ã®å€ãå ¥åããå Žåã¯ã CâããŒãæŒããŠããæ°åãå ¥ åããŠãã ããã æ°å€ã¯å°æ°ç¹ä»¥äžç¬¬2äœãŸã§å ¥å ã§ããŸãã 27 Equation 1 and Equation 2 èšç®åŒã®åç§° âš Equation èšç®åŒ âš Units èšç®çµæã®åäœïŒååïŒ âš âš If Yes Save Method If Yes Load and âš Enable Equation âš èšç®çµæã®åäœïŒåæ¯ïŒ âš âš Description âš âš 28 enter run ããŒãæŒããŠãªãã¡ã¬ã³ã¹ãåããç¶ããŠârun ããŒãæŒããŠãµã³ãã«ã枬å®ããŸãã èšç®çµæïŒæž¬å®åŸã«è¡šç€ºãããŸãïŒ Equation 1 Description Equation 2ã®å ¥åç»é¢ã« ã®ã¿è¡šç€ºãããŸãã Equation 2 Description Multiwaveã¢ãŒãã®å ¥åäŸ èšç®åŒ Cobalt ïŒg/lïŒ = ïŒïŒ12.26×A511ïŒâïŒ0.3×A720ïŒïŒ×100 Nickel ïŒg/lïŒ = ïŒïŒâ0.40×A511ïŒâïŒ27.41×A720ïŒïŒ×100 Set up âš 511 λ2 720 Integration Time 1s ⚠⚠λ1 âš âš æ°åãå ¥åãâãããã¯âããŒãæŒããŸãã enter No.of. λâs 2 ããŒã§éžæ Factors âš æ°åãå ¥åãâãããã¯âããŒãæŒããŸãã enter C 100 Î1 12.26 Î2 0.30 K3 -0.40 K4 27.41 Equation 1 and âš ãã€ãã¹ã®å€ãå ¥åããå Žåã¯ã CâããŒãæŒ ããŠããæ°åãå ¥åããŠãã ããã æ°å€ã¯å°æ°ç¹ä»¥äžç¬¬2äœãŸã§å ¥åã§ããŸãã Equation 2 Description Cobalt âš å€ãå ¥åãâããŒãæŒããŸãã âš ããŒã§éžæ âš enter ããŒã§ 1æåãã€éžæããâã§æ±ºå®ã ãŸããééããå Žå㯠C ã§æ¶å»ããŸãã âš âš âš âš âš ããŒãæŒããšãããŒããããçŸããŸãã 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