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Cell lysate¿¡¼­ His-tagged protein Á÷Á¢ Á¤Á¦

TALON (Cobalt) CellThru resin

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
635509
TALON CellThru Resin
°ü·ÃÇмú ±¸¸ÅÇϱâ
10 ml
292,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33126, x33127
Clontech
635510
TALON CellThru Resin
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 ml
2,014,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
635513
CellThru 10-ml Disposable Columns
°ü·ÃÇмú ±¸¸ÅÇϱâ
20 °³
286,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33126, x33127
Clontech
635651
HisTALON Buffer Set
°ü·ÃÇмú ±¸¸ÅÇϱâ
20 ȸ
472,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32713, x32714, x33126, x33127

  • ºü¸£°í °£ÆíÇÑ ÇÁ·ÎÅäÄÝ: crude cell lysate¿¡¼­ Á÷Á¢ Á¤Á¦
  • ¸·´Ü¹éÁú ¶Ç´Â multiprotein complexÀÇ Á¤Á¦¿¡ ÃÖÀû
  • Cobalt ÀÌ¿ÂÀ» ÀÌ¿ëÇÑ IMAC resin : Nickel resin º¸´Ù °í¼øµµ·Î his-tagged protein Á¤Á¦
Á¦Ç° ¼³¸í
TALON¢ç CellThru ResinÀº cell lysate, ÃÊÀ½ÆÄ Æļ⠻ùÇÃ, fermenter¿¡¼­ ȸ¼öÇÑ ¼¼Æ÷ µî¿¡¼­ his-tagged proteinÀ» Á¤Á¦ÇÒ ¼ö ÀÖ´Â IMAC resinÀÌ´Ù. ±âÁ¸ÀÇ TALON¢ç Metal Affinity Resin°ú °°ÀÌ µ¶ÀÚÀûÀÎ ¸®°£µå¸¦ °¡Áö°í ÀÖÁö¸¸ ´õ Å« beads (300-500§­)¸¦ »ç¿ëÇÏ¿© ¿ø½ÉºÐ¸® ¾øÀ̵µ ¼¼Æ÷ ÀÜÇع°ÀÌ resinÀ» Åë°úÇÒ ¼ö ÀÖ´Ù. ½Å¼ÓÇÑ 1 step °úÁ¤À¸·Î crude lysate¿¡¼­ his-tagged proteinÀ» Á÷Á¢ Á¤Á¦ÇϹǷÎ, ±âÁ¸ ¹æ¹ý º¸´Ù ´Ü¹éÁú ºÐÇØ °¡´É¼ºÀ» ³·Ãß°í ³ôÀº ¼öÀ²·Î ´Ü¹éÁúÀ» Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.
ÀϹÝÀûÀÎ resinÀ» »ç¿ëÇϸé cell lysate¸¦ column¿¡ Àû¿ëÇϱâ Àü¿¡ ¿ø½ÉºÐ¸® µîÀ» ÅëÇØ cell debris¸¦ Á¦°ÅÇÏ´Â °úÁ¤ÀÌ ÇÊ¿äÇѵ¥, ÀÌ ¶§ Á¤Á¦ÇÏ°íÀÚ ÇÏ´Â ¸· °áÇÕ ´Ü¹éÁú ¶Ç´Â ´Ü¹éÁú º¹ÇÕü°¡ ¼¼Æ÷ ÀÜ¿©¹°°ú ÇÔ²² Á¦°ÅµÇ´Â °æ¿ì°¡ ÀÖ´Ù. ¹Ý¸é, TALON¢ç CellThru ResinÀ» ÀÌ¿ëÇϸé cell lysateÀÇ ¿ø½ÉºÐ¸® °úÁ¤ ¾øÀÌ ±×´ë·Î resin¿¡ Àû¿ëÇÒ ¼ö ÀÖÀ¸¹Ç·Î, »ùÇà ¾È¿¡ Á¸ÀçÇÏ´Â ¸· °áÇÕ ´Ü¹éÁú ¶Ç´Â ´Ü¹éÁú º¹ÇÕü¸¦ ¼º°øÀûÀ¸·Î Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.
Å©±â°¡ Å« agarose beads·Î ±¸¼ºµÈ º» resinÀº ±¸¸ÛÀÌ ¸·È÷´Â °ÍÀ» ¸·±â À§ÇØ Áö¸§ÀÌ Å« ÀÏ¹Ý chromatography column¿¡ ÃæÁøÇؼ­ »ç¿ëÇϰųª, ¼¼Æ÷ ÀÜ¿©¹°À» ½±°Ô Åë°ú½Ãų ¼ö ÀÖµµ·Ï 90~130§­ÀÇ ÇÊÅ͸¦ ÀåÂøÇÏ¿© ÆǸŠÁßÀÎ CellThru-10§¢ Disposable Column°ú ÇÔ²² »ç¿ëÇÏ¸é µÈ´Ù.
TALON¢ç resinÀÇ Æ¯Â¡
TALON resin
TALON¢ç resinÀº cobalt¸¦ ÀÌ¿ëÇØ his-tagged proteinÀ» Á¤Á¦ÇÏ´Â immobilized metal affinity chromatography (IMAC) resinÀÌ´Ù. Cobalt´Â nickelº¸´Ù his-tagged protein¿¡ ´ëÇØ ³ôÀº ƯÀ̼ºÀ» ³ªÅ¸³»¹Ç·Î his-tagged proteinÀ» °í¼øµµ·Î Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.
Cobalt¸¦ Æ÷ÇÔÇÑ °áÇÕ ºÎÀ§
TALON¢ç Metal Affinity ResinÀº °áÇÕ ºÎÀ§¿¡ cobalt ÀÌ¿ÂÀ» Æ÷ÇÔÇÏ°í ÀÖ¾î, his-tagged protein¿¡ ´ëÇØ ¸Å¿ì ³ôÀº ƯÀ̼ºÀ» °¡Áö°í ÀÖ´Ù. Cobalt ÀÌ¿ÂÀ» Æ÷ÇÔÇÑ TALON¢ç °áÇÕºÎÀ§´Â histidineÀÇ À§Ä¡¿¡ ¸Å¿ì ±î´Ù·Î¿ö histidineÀÌ °ø°£ÀûÀ¸·Î ¿¬¼ÓÇØ Àְųª, °¡±îÀÌ À§Ä¡ÇÑ (6x HN tag, HAT tag µî) ´Ü¹éÁú¸¸ resin¿¡ °áÇÕÇÒ ¼ö ÀÖ´Ù. ¹Ý¸é nickel ±â¹ÝÀÇ resin (Ni-NTA resin µî)Àº ÀÌ·¯ÇÑ Æ¯ÀÌÀûÀÎ ¼­¿­ÀÌ ¾Æ´Ñ histidineÀÌ ºÒ±ÔÄ¢ÇÏ°Ô ¹èÄ¡µÈ ¼­¿­¿¡µµ °áÇÕÇϹǷÎ, »ó´ëÀûÀ¸·Î cobalt ±â¹ÝÀÇ TALON¢ç resinÀÌ nickel¿¡ ºñÇØ his-tagged proteinÀ» ƯÀ̼º ³ô°Ô Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.
±Ý¼ÓÀÌ¿Â Å»¶ôÀÌ ÀûÀº ±ÕÀÏÇÑ matrix ±¸Á¶
Cobalt resinÀº nickel resin¿¡ ºñÇØ ´õ ±ÕÀÏÇÑ ±¸Á¶¸¦ °¡Áø´Ù. TALON¢ç resinÀº 3Â÷ Æ÷Äϱ¸Á¶¸¦ Çü¼ºÇÏ¸ç °¢°¢ 3°³ÀÇ carboxyl ±×·ì°ú ÇϳªÀÇ Áú¼Ò¿øÀÚ°¡ cobalt ¾çÀ̿°ú °áÇÕÇÑ ±¸Á¶·Î, ÀÌ·¯ÇÑ ±¸Á¶´Â cobalt ÀÌ¿ÂÀÌ 2°³ÀÇ ÀÎÁ¢ÇÑ histidine Àܱâ¿Í °áÇյǵµ·Ï ÇÑ´Ù. µû¶ó¼­ cobalt ÀÌ¿ÂÀÌ ¸Å¿ì ´Ü´ÜÇÏ°Ô °áÇյǾî ÀÖ¾î resin¿¡¼­ ´©ÃâµÇÁö ¾Ê´Â´Ù. ¹Ý¸é nickel resinÀº nickel ÀÌ¿ÂÀÌ TALON¢ç ligand¿Í À¯»çÇÑ 3Â÷ Æ÷Äϱ¸Á¶³ª Æò¸é±¸Á¶, Áï 2°³ÀÇ ´Ù¸¥ ¹èÀ§±¸Á¶¸¦ Çü¼ºÇÒ ¼ö ÀÖ¾î TALON¢ç¿¡ ºñÇØ ±¸Á¶°¡ ´ú ±ÕÀÏÇÏ´Ù. Æò¸é ±¸Á¶¿¡¼­ nickel ÀÌ¿ÂÀº 2°³ÀÇ carboxyl ±×·ì°ú ÇϳªÀÇ Áú¼Ò¿øÀÚ¿¡ °áÇÕÇÏ°í ÀÖ¾î TALON¢ç¿¡ ºñÇØ °áÇÕÀÌ ¾àÇÏ°í nickel ÀÌ¿ÂÀ» °­ÇÏ°Ô À¯ÁöÇÒ ¼ö ¾ø¾î resin¿¡¼­ÀÇ ´©ÃâÀÌ ÀϾ´Ù. ±ÕÀÏÇÑ matrix¸¦ Çü¼ºÇÏ´Â TALON¢ç Metal Affinity ResinÀº ¿©·¯ Á¤Á¦ Á¶°Ç ÇÏ¿¡¼­ ³ôÀº °áÇÕ´É·ÂÀ» °¡Áö¹Ç·Î È®½ÇÇÏ°Ô his-tagged proteinÀ» Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.


±×¸². TALON¢ç CellThru ResinÀ» ÀÌ¿ëÇÑ ´Ü¹éÁú Á¤Á¦
E. coli BL21 À» TALON¢ç wash buffer¿¡¼­ sonication ÇÑ ÈÄ TALON¢ç CellThru columnÀ» ÀÌ¿ëÇÏ¿© 150 mM imidazole·Î Á¤Á¦ÇÏ¿´´Ù. M: molecular weight marker.
(Note: Membrane fraction¿¡ ³²¾ÆÀִ Ÿ°Ù ´Ü¹éÁúÀÇ ÀϺδ nonabsorbed material ±¸È¹¿¡¼­ È®ÀεÊ)


±×¸². TALON¢ç CellThru Resin & CellThru 10-§¢ Disposable Columns
º¸Á¸
TALON¢ç CellThru£º4¡É
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
°ü·ÃÁ¦Ç°
- Protein Á¤Á¦¸¦ À§ÇÑ Àü¿ë lysis buffer : xTractor¢â Buffer Kit
- His-tagged protein °í¹ßÇöÀ» À§ÇÑ cold shock vector system : pCold¢â vector
- Acetic acid ¾ø´Â °í°¨µµ ´Ü¹éÁú ¿°»ö½Ã¾à : TaKaRa CBB Protein Safe Stain
- ¹ü¿ëÀûÀÎ °ËÃâºÎÅÍ ÃÊ°í°¨µµ °ËÃâ±îÁö ´Ù¾çÇÑ Á¦Ç° ¶óÀξ÷ : HRP Substrate ½Ã¸®Áî
- ½Ç¿Â¿¡¼­ 30ºÐ ¿Ï·á : Western BLoT Stripping Buffer (Code T7135A)

Keyword :

His-tag À¶ÇÕ ´Ü¹éÁú Á¤Á¦
>
Spin columnŸÀÔÀ¸·Î ºü¸£°í °£ÆíÇÑ His-tag ´Ü..
Capturem¢â His-Tagged Purification
>
His-tagged protein °í¼øµµ Á¤Á¦
TALON (Cobalt) resin & column
>
ºü¸£°Ô ¼Ò¿ë·®ÀÇ his-tagged protein Á¤Á¦
TALON Magnetic Beads
>
FPLC purification - His-tagged protein
TALON (Cobalt) Superflow resin & cartridge
>
´ëÀå±ÕÀ¸·ÎºÎÅÍ His-tag À¶ÇÕ ´Ü¹éÁú ÃßÃâ¿¡,xT..
xTractor Buffer Kit
>
°í¼öÀ² Ni resin & FPLC purification
His60 (Nickel) Superflow resin & cartridge
>
High-throughput & microscale
His60 Ni Magnetic Beads
>
4¡É¿¡¼­ È°¼ºÀ» ¶ç´Â HN-tag¸¦ °®°í ÀÖ´Â ´Ü¹é..
HRV 3C Protease
>
Cell lysate¿¡¼­ His-tagged protein Á÷Á¢ Á¤Á¦
TALON (Cobalt) CellThru resin
>
Prepacked spin column
TALONSpin Column