![]() |
²úµØ | ÉϺ£ |
±£´æÌõ¼þ | 2-8¡æ |
Æ·ÅÆ | DUMABIO |
»õºÅ | DM-P5444 |
ÓÃ; | ¿ÆÑÐÓëʵÑé |
¼ì²â·½·¨ | Îü¸½²â¶¨·¨ |
±£ÖÊÆÚ | 6¸öÔÂ |
ÊÊÓ¦ÎïÖÖ | Öí |
¼ì²âÏÞ | Öí |
ÊýÁ¿ | 1000 |
°ü×°¹æ¸ñ | 96T/48T |
±ê¼ÇÎï | µçѯ |
Ñù±¾ | ѪÇå,ϸ°û,ϸ°ûÉÏÇåÒº¡¢ÌåÒºµÈ |
Ó¦Óà | ¿ÆÑÐÊÔ¼Á |
ÊÇ·ñ½ø¿Ú | ·ñ |
»õºÅ£ºYX-W-TPP ΢Á¿·¨ 100 ¹Ü/48 Ñù
º£ÔåÌÇÓÖ³ÆÎªÂ©Â«ÌÇ¡¢Þ¦ÌÇ£¬ÊÇÓÉÁ½¸öÆÏÌÑÌÇ·Ö×Ó×é³ÉµÄÒ»¸ö·Ç»¹ÔÐÔË«ÌÇ£¬Äܹ»ÔÚ¸ßΡ¢¸ßº®¡¢¸ßÉøÍ¸Ñ¹¼°¸ÉÔïʧˮµÈ¶ñÁÓ»·¾³Ìõ¼þÏÂÔÚϸ°û±íÃæÐγɶÀÌØµÄ±£»¤Ä¤£¬ÓÐЧµØ±£»¤ÉúÎï·Ö×ӽṹ²»±»ÆÆ»µ£¬´Ó¶øÎ¬³ÖÉúÃüÌåµÄÉúÃü¹ý³ÌºÍÉúÎïÌØÕ÷¡£
Ö²ÎïÌåÄÚÖ÷ÒªÒÔÆÏÌÑÌÇΪµ×ÎïºÏ³Éº£ÔåÌÇ£¬º£ÔåÌÇ-6-Á×Ëáõ¥Ã¸ £¨trehalose 6-phosphate phosphatase£¬TPP£©ÊǸÃ;¾¶Öк£ÔåÌǺϳɹؼüø֮һ£¬¸Ã·´Ó¦Ê×ÏÈÔÚº£ÔåÌÇ-6-Á×ËáºÏ³Éø
£¨trehalose-6-phosphate synthase£¬TPS£©µÄ×÷ÓÃÏ£¬´ß»¯ÄòÜÕ¶þÁ×ËáÆÏÌÑÌÇ£¨UDPG£©ºÍ 6-
Á×ËáÆÏÌÑÌÇÉú³ÉÖмä²úÎï6-Á×ËảÔåÌÇ£¬ÔÙÔÚTPPµÄ×÷ÓÃÏÂÈ¥Á×Ëữ£¬Éú³Éº£ÔåÌÇ¡£
TPP ´ß»¯º£ÔåÌÇ-6-Á×Ëá²úÉúº£ÔåÌÇ£¬Í¬Ê±ÊͷųöÁ×Ëá¸ù£¬Ê¹ÓÃîâÌ࿹±ÈÉ«·¨²â¶¨¡£
ø±êÒÇ¡¢Ë®Ô¡¹ø¡¢¿Éµ÷Ê½ÒÆÒºÆ÷¡¢96 ¿×°å¡¢Ñв§¡¢±ùºÍÕôÁóË®¡£
ÌáȡҺ£ºÒºÌå 100mL¡Á1 Æ¿£¬4¡æ±£´æ£» ÊÔ¼ÁÒ»£ºÒºÌå 12mL¡Á1 Æ¿£¬4¡æ±£´æ£» ÊÔ¼Á¶þ£ºÒºÌå 6mL¡Á1 Ö§£¬4¡æ±£´æ¡£
ÊÔ¼ÁÈý£º ·Û¼Á¡Á1 Ö§£¬4¡æ±Ü¹â±£´æ¡£ÁÙÓÃǰ¼Ó 2mL ÕôÁóË®Èܽ⡣Óò»ÍêµÄÊÔ¼Á 4¡æ±£´æ¡£ÊÔ¼ÁËÄ£º ·Û¼Á¡Á3 Ö§£¬4¡æ±Ü¹â±£´æ¡£ÁÙÓÃǰ¼Ó 1mL ÕôÁóË®Èܽ⡣ÏÖÅäÏÖÓá£
1¡¢Ï¸¾ú»òϸ°ûµÄ´¦Àí£ºÊÕ¼¯Ï¸¾ú»òϸ°ûµ½ÀëÐĹÜÄÚ£¬ÀëÐĺóÆúÉÏÇ壻°´ÕÕϸ¾ú»òϸ°ûÊýÁ¿
£¨104 ¸ö£©£ºÌáȡҺÌå»ý£¨mL£©Îª 500~1000£º1 µÄ±ÈÀý£¨½¨Òé 500 Íòϸ¾ú»òϸ°û¼ÓÈë 1mL ÌáȡҺ£©£¬³¬Éù²¨ÆÆËéϸ¾ú»òϸ°û£¨±ùÔ¡£¬¹¦ÂÊ 20£¥»ò 200W£¬³¬Éù 3S£¬¼ä¸ô 10S£¬Öظ´30 ´Î£©£»8000g£¬4¡æÀëÐÄ 10min£¬È¡ÉÏÇ壬ÖñùÉÏ´ý²â¡£
2¡¢×éÖ¯µÄ´¦Àí£º°´ÕÕ×éÖ¯ÖÊÁ¿£¨g£©£ºÌáȡҺÌå»ý(mL)Ϊ 1£º5~10 µÄ±ÈÀý£¨½¨Òé³ÆÈ¡Ô¼ 0.1g ×éÖ¯£¬¼ÓÈë 1mL ÌáȡҺ£©£¬±ùÔ¡ÖÐÔȽ¬¡£8000g £¬4¡æÀëÐÄ 10min£¬È¡ÉÏÇ壬ÖñùÉÏ´ý²â¡£²â¶¨²½Öè
1¡¢ÔÚEP ¹ÜÖмÓÈëÈçÏÂÊÔ¼Á
ÊÔ¼ÁÃû³Æ£¨¦ÌL£© |
¶ÔÕÕ¹Ü |
²â¶¨¹Ü |
Ñù±¾ |
|
100 |
95¡æË®Ô¡Ãð»îÑù±¾ |
100 |
|
ÊÔ¼ÁÒ» |
100 |
100 |
»ìÔÈ£¬37¡æ·´Ó¦ 30min£¬95¡æË®Ô¡ 5min ÖÕÖ¹·´Ó¦£¬ÀäÈ´ÖÁÊÒΡ£10000g 4¡æÀëÐÄ 5min£¬È¡·´Ó¦Òº´ý²â¡£
2¡¢ÏÔɫҺµÄÅäÖÆ£¨¿É²â 50 ¹Ü£©£ºÈ¡ EP ¹ÜÒ»Ö§£¬¼ÓÈë 660¦ÌL ÊÔ¼Á¶þ£¬ÔÙ¼ÓÈë 100¦ÌL ÊÔ¼ÁÈý£¬ ³ä·Ö»ìÔȺó£¬ÔÙ¼ÓÈë 240¦ÌL ÊÔ¼ÁËÄ£¬³ä·Ö»ìÔÈ´ýÓã»ÅäºÃµÄÏÔɫҺӦΪ»ÆÉ«£¬Èô±äÀ¶ÔòΪÁ×ÎÛȾ£»ÏÔɫҺ±ØÐëÏÖÅäÏÖÓã»ÈôÒ»´ÎÐԲⲻÁË 50 ¸öÑù£¬¿É°´±ÈÀýËõС¸÷ÊÔ¼ÁÌå»ý¡£
3¡¢ÔÚ 96 ¿×°åÖмÓÈëÈçÏÂÊÔ¼Á
|
¶ÔÕÕ¹Ü |
²â¶¨¹Ü |
²â¶¨·´Ó¦Òº£¨¦ÌL£© |
|
40 |
¶ÔÕÕ·´Ó¦Òº£¨¦ÌL£© |
40 |
|
ÏÔɫҺ£¨¦ÌL£© |
20 |
20 |
ÕôÁóË®£¨¦ÌL£© |
140 |
140 |
²â¶¨ 660nm ´¦Îü¹âÖµA£¬·Ö±ð¼ÇΪA ¶ÔÕÕºÍ A ²â¶¨£¬¡÷A=A ²â¶¨-A ¶ÔÕÕ¡£
±ê×¼Ìõ¼þϱê×¼ÇúÏßΪ y = 1.9403x - 0.0036£¬R2 = 0.9985£»ÆäÖÐ x Ϊ±ê׼ƷŨ¶È£¬¦Ìmol/mL£¬
y ΪÎü¹âÖµ¡÷A¡£
1¡¢°´ÕÕµ°°×Ũ¶È¼ÆËã
µ¥Î»µÄ¶¨Ò壺ÿ mg ×éÖ¯µ°°×ÿ·ÖÖÓ´ß»¯²úÉú 1nmol ÎÞ»úÁ×¶¨ÒåΪһ¸öø»îÁ¦µ¥Î»¡£
TPP »îÁ¦( nmol/min/mg prot)=£¨¦¤A+0.0036£©¡Â1.9403¡ÁV ·´×ܡ£¨V Ñù¡ÁCpr £©¡ÂT¡Á 1000
=34.35¡Á£¨¦¤A+0.0036£©¡ÂCpr
2¡¢°´Ñù±¾ÏÊÖØ¼ÆËã
µ¥Î»µÄ¶¨Ò壺ÿg ×é֯ÿ·ÖÖÓ´ß»¯²úÉú 1nmol ÎÞ»úÁ×¶¨ÒåΪһ¸öø»îÁ¦µ¥Î»¡£
TPP »îÁ¦(nmol/min/g ÏÊÖØ)=£¨¦¤A+0.0036£©¡Â1.9403¡ÁV ·´×ܡ£¨W¡Á V Ñù¡ÂV Ñù×Ü£©¡ÂT¡Á 1000
=34.35¡Á£¨¦¤A+0.0036£©¡ÂW
3¡¢°´Ï¸¾ú»òϸ°ûÃܶȼÆËã
µ¥Î»µÄ¶¨Ò壺ÿ 1 Íò¸öϸ¾ú»òϸ°ûÿ·ÖÖÓ´ß»¯²úÉú 1nmol ÎÞ»úÁ×¶¨ÒåΪһ¸öø»îÁ¦µ¥Î»¡£
TPP »îÁ¦(nmol/min/104 cell)=£¨¦¤A+0.0036£©¡Â1.9403¡ÁV ·´×ܡ£¨V Ñù¡Áϸ°ûÊýÁ¿£©¡ÂT¡Á 1000
=0.069¡Á£¨¦¤A+0.0036£©
V ·´×Ü£º·´Ó¦Ìåϵ×ÜÌå»ý£¬0.2mL£»V Ñù£º¼ÓÈëÑù±¾Ìå»ý£¬0.1 mL£»V Ñù×Ü£º¼ÓÈëÌáȡҺÌå»ý£¬1 mL£»T£º·´Ó¦Ê±¼ä£¬30 min£»Cpr£ºÑù±¾µ°°×ÖÊŨ¶È£¬mg/mL£»W£ºÑù±¾ÖÊÁ¿£»Ï¸°ûÊýÁ¿£¬500 Íò£»1000£¬¦Ìmol µ½ nmol µÄ»»ËãϵÊý¡£