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DIGE - ¼ºñ½º ÀýÂ÷¿Í ¼ö°¡
1. DIGE ¼ºñ½º ¼ö°¡ : IPG strip 24cm ´ç 3,000,000¿ø ( 3 DIGE gel + 1 prep gel + Image Analysis)
2. ¼ºñ½º ÀýÂ÷
<Sample preparation-protein Á¤·®-CyDye labeling-IEF-2DE-image scanning-Statistical analysis>
- »ùÇÃÀº ÃÖ¼Ò 1.0 mg Á¤µµ ¼Ò¿äµË´Ï´Ù. (ºñ±³´ë»óÀÌ 2°³ÀÏ °æ¿ì 75ug/DIGE gel, 500ug/prep gelÀÌ ÇÊ¿ä)
- tissue ÀÎ °æ¿ì ±×´ë·Î ³Ãµ¿»óÅ·Πº¸³»ÁÖ½Ã¸é µË´Ï´Ù.
- cell ÀÎ °æ¿ì 150mm dish ÀÌ»ó ÇÊ¿äÇÕ´Ï´Ù. 1x PBS > 1/2x PBS > 1/4x PBS ·Î ¼øÂ÷ÀûÀ¸·Î washing ÇϽðí centrifugation ÇϽŠÈÄ pellet ¸¸ º¸³»½Ã¸é µË´Ï´Ù.
- fluid, sereum ÀÎ °æ¿ì´Â »ùÇà ÀÚü¸¦ Á¤·®Çؼ 1mg µÇ°Ô º¸³»ÁÖ½Ã¸é µË´Ï´Ù.
- 1D (Isoelectric focusion) -> 2DE -> Scanning ±îÁö ¼Ò¿äµÇ´Â ½Ã°£Àº 7ÀÏÀÔ´Ï´Ù.
- °á°ú º¸°í¼ ¾ç½Ä : SpotÀº p<0.05 ·Î Áõ°¨µÈ °ÍÀ¸·Î selection Çϸç, ºÐ¼®µÈ Image´Â PPTÆÄÀÏ·Î Àüü À̹ÌÁö ¿Ü, Â÷ÀÌ ³ª´Â spotsÀ» È®ÀÎÀÌ ½±°Ô È®´ëÇÏ¿© º¸³»µå¸®¸ç, ½ÇÇè design ¹× °úÁ¤À» Á¤¸®ÇÑ ÆÄÀÏÀ» ÇÔ²² º¸³»µå¸³´Ï´Ù. ºÐ¼® ÀÌÈÄ ¿øÇϽô ºÐ¼®Á¤º¸ ( raw data, 3D À̹ÌÁö) µµ ÇÊ¿äÇÏ½Ã¸é º¸³»µå¸³´Ï´Ù.
- ¼ºñ½º ½ÅûÀ» ÇϽñâ Àü, ¹Ýµå½Ã ¿¬±¸¿ø (ÀÌÀº¿µ ¹Ú»ç :02-2123-6625,
)°ú ÇØ´ç ¼ºñ½º¿¡ ´ëÇØ¼ ²À ¹®ÀÇÇϽŠÈÄ, ¼ºñ½º ½ÅûÀ» ÇϽñ⠹ٶø´Ï´Ù.
- »ùÇà º¸³»½Ç ÁÖ¼Ò : ¼¿ï ¼´ë¹®±¸ ½ÅÃ̵¿ 134 ¿¬¼¼´ëÇб³ »êÇÐÇùµ¿°ü 421È£.
- ºÐ¼® ¼ºñ½º ¿ä±ÝÀº ¿¬¼¼´ëÇб³ °èÁ·ΠÀԱݽÃÄÑ ÁÖ½Ã¸é µË´Ï´Ù. ¿¬¼¼´ëÇб³ ¸íÀÇÀÇ °è»ê¼°¡ ¹ßºÎµÇ¸ç, ¼¼±Ý°è»ê¼´Â ¹ß±ÞÀÌ ¾ÈµÇ°í, Ä«µå °áÀçµµ ¾ÈµË´Ï´Ù. °áÀç°èÁ¾ȳ»´Â ¸ÞÀÏ·Î µå¸³´Ï´Ù.
3. ºÐ¼®¼ºñ½º ¼Ò¿ä½Ã°£
| 2-D DIGE 7 working day |
| Sample preparation (sonication, ultracentrifuge)- protein Á¤·® |
5½Ã°£ |
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| rehydration |
16~18½Ã°£ |
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| Focusing |
37½Ã°£ |
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| gel running |
6-10½Ã°£ |
| DIGE gel scanning |
2½Ã°£ / gel |
| Prep gel Coomassie staining |
12½Ã°£ |
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| Prep gel destaining |
4-5½Ã°£ |
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| Prep gel scanning |
1½Ã°£ |
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Image analysis (DeCyder software) Spot detection, matching, editing, reporting
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2ÀÏ |
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ÃÑ ¼Ò¿ä ±â°£
Pre-test 2DE -> DIGE -> Image analysis / reporting : 4ÁÖ
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