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Home > ÀüÁ¦Ç°º¸±â > ´Ü¹éÁú ¹ßÇö ¡¤ Á¤Á¦ > ´ëÀå±Õ ´Ü¹éÁú ¹ßÇö > HAT ´Ü¹éÁú ¹ßÇö£¦ Á¤Á¦ ½Ã½ºÅÛ

HAT ´Ü¹éÁú ¹ßÇö£¦ Á¤Á¦ ½Ã½ºÅÛ

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
631205
HAT Protein Expression & Purification System
°ü·ÃÇмú ±¸¸ÅÇϱâ
1 Set
(Package)
1,420,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33377, x32699


  • Áß¼º pH »óÅ¿¡¼­ ´Ü¹éÁú Á¤Á¦
  • Native ȤÀº denaturation »óÅ¿¡¼­ »ç¿ë
  • ´õ ³ôÀº ´Ü¹éÁú ¿ëÇصµ
  • TALON IMAC resinÀ» ÀÌ¿ëÇÑ Á¤Á¦¿¡ ÃÖÀû
Á¦Ç°¼³¸í
HAT Protein Expression and Purification System (ƯÇã Ãâ¿ø Áß)À» ÀÌ¿ëÇϸé, ´Ü¹éÁú ¹ßÇö°ú Á¤Á¦¸¦ º¸´Ù ½±°í È¿À²ÀûÀ¸·Î ÁøÇàÇÒ ¼ö ÀÖ´Ù. E.coli¿¡ HAT vector°¡ µµÀԵǸé, HAT tag¸¦ Æ÷ÇÔÇÑ ¸ñÀû ´Ü¹éÁúÀ» »ý»êÇÒ ¼ö ÀÖ°í, »ý»êµÈ ¸ñÀû ´Ü¹éÁúÀº »ýü¿Í µ¿ÀÏÇÑ Áß¼º pH »óÅ¿¡¼­ Á¤Á¦ÇÑ´Ù. HAT (histidine affinity tag) tag´Â õ¿¬ polyhistidine peptide·Î, inclusion body¸¦ Çü¼ºÇÏÁö ¾Ê´Â °ÍÀÌ ÀåÁ¡ÀÌ´Ù. ¶ÇÇÑ, HAT tag´Â 6 x Hisº¸´Ù ±æÀÌ°¡ ±æ¾î, ¸ñÀû ´Ü¹éÁúÀÌ °íºÐÀÚÀÎ °æ¿ì °íºÐÀڴܹéÁú ±¸Á¶ ¹Ù±ùÂÊ¿¡ À§Ä¡ÇÏ°Ô µÈ´Ù. µû¶ó¼­, Tag°¡ ´Ü¹éÁú ±¸Á¶ ³»ºÎ¿¡ À§Ä¡ÇÒ °æ¿ì¿¡ ºñÇØ histidineÀÌ Á¤Á¦ resin°ú ´õ ½±°Ô °áÇÕÇÏ¿©, ¸ñÀû ´Ü¹éÁúÀÇ Á¤Á¦°¡ ¿ëÀÌÇÏ´Ù. ½Ã½ºÅÛ ³» Æ÷ÇԵǾî ÀÖ´Â TALON resinÀ» ÀÌ¿ëÇϸé, º¯¼º ¹× ºñº¯¼º Á¶°Ç ¸ðµÎ¿¡¼­ ¸ñÀû ´Ü¹éÁúÀ» ½±°Ô Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.


±×¸² 1. TALON SuperflowÀ» ÀÌ¿ëÇÑ HAT-Tag À¶ÇÕ ´Ü¹éÁúÀÇ FPLC Á¤Á¦
E. coli ´Â 50 mM sodium phosphate, 0.3 M sodium chloride, 5 mM imidazoleÀ» Æ÷ÇÔÇÏ´Â ½Ã¾à (pH 7.0) À¸·Î ÃßÃâÇÏ¿©, 150 mM imidazole ¿ë¾×À¸·Î ´Ü¹éÁúÀ» ȸ¼öÇÏ¿´´Ù.
(A) ¼¼Æ÷ ¿ëÇØ ÈÄ, TALON Superflow columnÀ¸·Î ´Ü¹éÁú Á¤Á¦
(B) SDS-PAGE¸¦ ÀÌ¿ëÇÑ Á¤Á¦ È®ÀÎ (M: Molecular weight marker)

Ç¥ 1. Polyhistidine tag sequence
tag

Amino acid sequence

6 x His

His -His -His -His -His -His

6 x HN

His -Asn -His -Asn -His -Asn -His -Asn -His -Asn -His -Asn

HAT

Lys -Asp -His -Leu -lle -His -Asn -Val -His -Lys -Glu -Glu -His -Ala -His -Asn -Lys



Ç¥ 2. HAT Protein Expression & Purification System (Code 631205)ÀÇ Æ¯Â¡°ú ÀåÁ¡
Ư¡

ÀåÁ¡

±ä ±æÀÌÀÇ tag

°íºÐÀÚ ´Ü¹éÁú¿¡ ÃÖÀû

Tag ³» ±ÕµîÇÑ ÀüÇÏ ºÐÆ÷

³ôÀº ¼ö¿ë¼º

õ¿¬ À¯·¡ÀÇ tag sequence

³·Àº ¼¼Æ÷ µ¶¼º

»ýü¿Í µ¿ÀÏÇÑ pH¿¡¼­ Á¤Á¦

¸ñÀû ´Ü¹éÁú ±¸Á¶ À¯Áö



±×¸² 2. pHAT 10/11/12ÀÇ vector map
HAT system´Â ¸ñÀû ´Ü¹éÁú ¼­¿­À» cloningÇÒ ¼ö ÀÖ´Â ¼¼ Á¾·ùÀÇ vector (pHAT10/11/12)°¡ Æ÷ÇԵǾî ÀÖÀ¸¸ç, °¢°¢ µ¿ÀÏÇÑ ±¸Á¶·Î MCS¿¡ ¼­·Î ´Ù¸¥ reading frameÀ» °¡Áø´Ù. ¶ÇÇÑ vector ³» Á¤Á¦µÈ ´Ü¹éÁú¿¡¼­ HAT sequence¸¦ Á¦°ÅÇÒ ¼ö ÀÖ´Â enterokinase (EK) cleavage site¸¦ Æ÷ÇÔÇÑ´Ù. Á¦ÇÑÈ¿¼Ò¸¦ ÀÌ¿ëÇϸé EK site¸¦ Æ÷ÇÔÇÑ HAT sequence ȤÀº ´Üµ¶À¸·Î ´Ù¸¥ vector¿¡ cloningÇÒ ¼ö ÀÖ´Ù.
±¸¼ºÇ° ¹× º¸Á¸ (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶ ¹Ù¶ø´Ï´Ù)

Keyword : ´Ü¹éÁú ¹ßÇö,´Ü¹éÁú Á¤Á¦,Ecoli,´ëÀå±Õ ¹ßÇö,IMAC,metal affinity,HAT,histidine affinity tag,His-tag

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