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1. Free Flow Electrophoresis ¼­ºñ½º ¼ö°¡

FFE (0.7 ml 70 fraction) + 1 NuPAGE service charge

Protocol
MP
MGP
UMP
High HP
FFE + 1 NuPAGE
Mannitol 4.56
Prolyte 25
Mannitol 4.56
Glycerol 25
Prolyte 25
Urea 8
Mannitol 4.0
Prolyte 25
HPMC 0.2
Prolyte16.6
Sample No. 1
260,000
270,000
270,000
230,000
2
440,000
450,000
450,000
390,000
3
620,000
630,000
630,000
550,000
4
800,000
810,000
810,000
710,000
5
980,000
910,000
910,000
870,000

Protocol
High HGP
High UHGP
Low HGP
Low UHGP
FFE + 1 NuPAGE
HPMC 0.2
Glycerol 25
Prolyte16.6
Urea 8
HPMC 0.12
Glycerol 14.5
Prolyte 14.5
HPMC 0.05
Glycerol 25
Prolyte16.6
Urea 8
HPMC 0.03
Glycerol 14.5
Prolyte14.5
Sample No. 1
230,000
230,000
230,000
230,000
2
400,000
400,000
390,000
400,000
3
570,000
570,000
550,000
570,000
4
740,000
740,000
710,000
740,000
5
910,000
910,000
870,000
910,000


2. Free Flow Electrophoresis_¼­ºñ½º ¾È³»

  1. »ùÇÃÀº 1.0~6.0 mg proteinÀ¸·Î °ÇÁ¶µÇ¾îÀÖ´Â »óÅÂÀ̰ųª low salt buffer ¿¡ ³ì¿©ÀÖ´Â »óÅÂÀ̾î¾ß ÇÏ¸ç »ùÇà ´ç Àüü volumeÀÌ 0.1 mlÀ» ³ÑÁö ¾Êµµ·Ï ÇØ¾ß ÇÕ´Ï´Ù.

  2. ÃÖ´ë 5°³ sample ±îÁö Çѹø¿¡ running °¡´ÉÇϸç, sampleÀÌ ¿¹¸¦ µé¾î 7°³Àΰæ¿ì¿¡´Â 2°³, 5°³·Î ³ª´² ¸Ã±â¼Å¾ß ÇÕ´Ï´Ù. °°Àº Á¾·ùÀÇ »ùÇõµ 0.1 ml ¶Ç´Â Àüü ´Ü¹éÁú·®ÀÌ 6 mg À» ³ÑÀ» °æ¿ì ¿¡´Â µÎ °³ÀÇ »ùÇ÷Π°£Áֵ˴ϴÙ. ÀÚ¼¼ÇÑ Á¶°ÇÀº »ùÇà ÀÇ·Ú Àü¿¡ ¹®ÀÇÇϽñ⠹ٶø´Ï´Ù.

  3. Serum Àº Àü ó¸® ¾øÀÌ ±×³É °¡Á®¿À½Ã¸é µË´Ï´Ù.

  4. ¸ðµç »ùÇÃÀº urea °¡ ÀÖÀ» ¶§¿Í ¾øÀ» ¶§ µÑ ´Ù running °¡´ÉÇÕ´Ï´Ù. Running protocol Àº urea °¡ µé¾î°¡´Â Á¶°Ç¿¡¼­´Â Low UHGPÀ» ±ÇÀåÇÕ´Ï´Ù. Urea °¡ µé¾î°¡Áö ¾Ê´Â Á¶°Ç¿¡¼­´Â High HGP¸¦ »ç¿ëÇÏ´Â °ÍÀÌ ÁÁ°í, glycerol ÀÌ ÈÄ󸮿¡ À־ ¹®Á¦°¡ µÇ´Â °æ¿ì protocol MP¸¦ »ç¿ëÇÏ°Ô µÇ³ª, High HGP ¿¡ ºñÇØ separation È¿À²Àº Á» ¶³¾îÁý´Ï´Ù.

  5. Fraction Àº ÃÖ¼Ò 14°³¿¡¼­ ÃÖ´ë 70°³±îÁö ¾òÀ» ¼ö ÀÖÀ¸¸ç 14, 28, 42, 56, 70 °³ Áß¿¡¼­ ¼±ÅÃÇÒ ¼ö ÀÖÀ¸³ª, 56 ¶Ç´Â 70°³¸¦ ±ÇÀåÇÕ´Ï´Ù.

  6. Focusing range ´Â À̷лó pH 2.5 ~ 11±îÁö °¡´ÉÇϰí, acidic ¶Ç´Â basic region ¸¸ ¼±º°ÇÏ¿© running ÇÒ ¼ö ÀÖÀ¸³ª, pH 3 ~ 10 range Á¶°ÇÀ» ±ÇÀåÇÕ´Ï´Ù.

  7. Fraction Àº well ´ç 0.6~0.7 ml Á¤µµ ¾ò¾îÁö°í ±× ³óµµ´Â runningÇÑ proteinÀÌ ¸ðµç well·Î °ñ°í·ç ÆÛÁ³´Ù°í °è»êÇÒ ¶§ 0.04~0.08 mg/ml ÀÔ´Ï´Ù. Protein ȸ¼öÀ²Àº ~90% Á¤µµÀÔ´Ï´Ù.

  8. FractionÀº deep well plate(1ml) ¿¡ ´ã¾Æ¼­ µå¸®°í ±× Áß ¸î °³ fraction À» 4-12% Bis-Tris 12 well small gels ÇÑ Àå¿¡ °É¾î¼­ ÷ºÎÇÏ¿© µå¸³´Ï´Ù.

  9. Sample preparation °úÁ¤Àº Á¦½ÃµÈ ºñ¿ë¿¡ Æ÷ÇԵǾî ÀÖÁö ¾Ê½À´Ï´Ù.

  10. Sample preparation °úÁ¤¿¡¼­ÀÇ buffer Á¶¼º (urea, salt ³óµµ, detergent)Àº ÀÇ·Ú Àü¿¡ ¹Ýµå½Ã ¹®ÀÇÇϽñ⠹ٶø´Ï´Ù.

  11. Àý´ë ¾à¼Ó ½Ã°£: ½Åû¾ç½Ä ¶Ç´Â »ùÇà Á¢¼ö ÈÄ, Business Day 3 weeks

  12. ¼­ºñ½º ÀýÂ÷, °á°ú º¸°í¼­ ¾ç½Ä µî ¾È³»ÀÚ·á ( FFE_¼­ºñ½º¾È³»_°ø°í¿ë.ppt)

  13. ¼­ºñ½º ½ÅûÀ» ÇϽñâ Àü, ¹Ýµå½Ã ¿¬±¸¿ø (ÀÌÀº¿µ :02-2123-6625, )°ú ÇØ´ç ¼­ºñ½º¿¡ ´ëÇØ¼­ ²À ¹®ÀÇÇϽŠÈÄ, ¼­ºñ½º ½ÅûÀ» ÇϽñ⠹ٶø´Ï´Ù.

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

  15. 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.




2008 © Yonsei Proteome Research Center #422, YPRC/BPRC, Industry-University Research Center, Yonsei Univ., Seodaemun-gu, Seoul, Korea, 120-749
Tel: +82-2-2123-3279, Fax: +82-2-393-6589