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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»
- »ùÇÃÀº 1.0~6.0 mg proteinÀ¸·Î °ÇÁ¶µÇ¾îÀÖ´Â »óÅÂÀ̰ųª low salt buffer ¿¡ ³ì¿©ÀÖ´Â »óÅÂÀ̾î¾ß ÇÏ¸ç »ùÇÃ
´ç Àüü volumeÀÌ 0.1 mlÀ» ³ÑÁö ¾Êµµ·Ï ÇØ¾ß ÇÕ´Ï´Ù.
- 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.
- Serum Àº Àü ó¸® ¾øÀÌ ±×³É °¡Á®¿À½Ã¸é µË´Ï´Ù.
- ¸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󸮿¡ ÀÖ¾î¼ ¹®Á¦°¡ µÇ´Â °æ¿ì protocol MP¸¦ »ç¿ëÇÏ°Ô µÇ³ª, High HGP ¿¡ ºñÇØ separation È¿À²Àº Á»
¶³¾îÁý´Ï´Ù.
- 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.
- 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.
- 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.
- 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.
- Sample preparation °úÁ¤Àº Á¦½ÃµÈ ºñ¿ë¿¡ Æ÷ÇԵǾî ÀÖÁö ¾Ê½À´Ï´Ù.
- Sample preparation °úÁ¤¿¡¼ÀÇ buffer Á¶¼º (urea, salt ³óµµ, detergent)Àº ÀÇ·Ú Àü¿¡ ¹Ýµå½Ã
¹®ÀÇÇϽñ⠹ٶø´Ï´Ù.
- Àý´ë ¾à¼Ó ½Ã°£: ½Åû¾ç½Ä ¶Ç´Â »ùÇà Á¢¼ö ÈÄ, Business Day 3 weeks
- ¼ºñ½º ÀýÂ÷, °á°ú º¸°í¼ ¾ç½Ä µî ¾È³»ÀÚ·á (
FFE_¼ºñ½º¾È³»_°ø°í¿ë.ppt)
- 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.
- »ùÇà º¸³»½Ç ÁÖ¼Ò : ¼¿ï ¼´ë¹®±¸ ½ÅÃ̵¿ ¿¬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.
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