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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>

  1. »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)

  2. tissue ÀÎ °æ¿ì ±×´ë·Î ³Ãµ¿»óÅ·Πº¸³»ÁÖ½Ã¸é µË´Ï´Ù.

  3. 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.

  4. fluid, sereum ÀÎ °æ¿ì´Â »ùÇà ÀÚü¸¦ Á¤·®Çؼ­ 1mg µÇ°Ô º¸³»ÁÖ½Ã¸é µË´Ï´Ù.

  5. 1D (Isoelectric focusion) -> 2DE -> Scanning ±îÁö ¼Ò¿äµÇ´Â ½Ã°£Àº 7ÀÏÀÔ´Ï´Ù.

  6. °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.

  7. 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.

  8. »ùÇà º¸³»½Ç ÁÖ¼Ò : ¼­¿ï ¼­´ë¹®±¸ ½ÅÃ̵¿ 134 ¿¬¼¼´ëÇб³ »êÇÐÇùµ¿°ü 421È£.

  9. 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°£
    rehydration
    16~18½Ã°£
    Focusing
    37½Ã°£
    gel running
    6-10½Ã°£
    DIGE gel scanning
    2½Ã°£ / gel
    Prep gel Coomassie staining
    12½Ã°£
    Prep gel destaining
    4-5½Ã°£
    Prep gel scanning
    1½Ã°£
    Image analysis (DeCyder software)
    Spot detection, matching, editing, reporting
    2ÀÏ

    ÃÑ ¼Ò¿ä ±â°£
    Pre-test 2DE -> DIGE -> Image analysis / reporting : 4ÁÖ

Dige



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Tel: +82-2-2123-3279, Fax: +82-2-393-6589