Myc-Tag & HA-Tag Vectors
- Æ÷À¯·ù ¼¼Æ÷¿¡¼ myc-tagged ¶Ç´Â HA-tagged proteinsÀÇ Co-immunoprecipitation¿¡ Àû¿ë
- Áö¼ÓÀûÀÌ°í ³ôÀº ´Ü¹éÁú ¹ßÇö
- Æ÷À¯·ù ¼¼Æ÷¿¡¼ yeast two-hybrid interactions È®Àο¡ Àû¿ë
- Western blotÀ» ÅëÇØ ¼±¸íÇÑ band È®ÀÎ °¡´É
c-MycÀ̳ª hemagglutinin (HA) epitope tagÀ¸·Î Ç¥½ÄµÈ ´Ü¹éÁú Æ÷À¯·ù ¹ßÇö º¤ÅÍÀÌ´Ù. c-Myc¿Í HA´Â ¸é¿ª¹ÝÀÀ¿ëÀ¸·Î ƯȵǾî ÀÖ¾î co-immunoprecipitation ¿¬±¸ µî¿¡
ÀÌ»óÀûÀ̸ç western blotÀ¸·Î ½±°Ô °ËÃâ ÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ c-Myc¿Í HA´Â ÀÛÀº tag·Î ¸ñÀû À¯ÀüÀÚÀÇ ±â´É¿¡ ¿µÇâÀ» ÁÖÁö ¾Ê´Â´Ù.
pCMV vector´Â CMV promoter¸¦ ÀÌ¿ëÇØ ´Ù¾çÇÑ Æ÷À¯·ù ¼¼Æ÷¿¡¼ Áö¼ÓÀûÀÌ°í ³ôÀº ¹ßÇöÀ» º¸ÀδÙ. MycÀ¸·Î Ç¥ÁöµÈ ¸ñÀû ´Ü¹éÁúÀº western blotÀ̳ª anti-myc antibody¸¦
»ç¿ëÇÏ¿© fluorescence microscopy·Î °ËÃâÇÏ´Â ¹æ¹ýÀÌ ÀÖ´Ù.
pCMV-Myc & pCMV-HA Vector Set´Â c-Myc Monoclonal Antibody¿Í HA-Tag Polyclonal Antibody¸¦ Æ÷ÇÔÇÏ°í ÀÖÀ¸¸ç ÀÌ Ç×ü¸¦ ÀÌ¿ëÇϸé Western blot, ELISA, ¶Ç´Â
¸é¿ª¿°»ö¹ý immunostaining assays¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ insertÀÇ reading frameÀÌ ¿Ã¹Ù¸¥Áö ¿©ºÎ¸¦ È®ÀÎÇÒ ¼ö ÀÖ´Â sequencing primer°¡ Æ÷ÇԵǾî ÀÖ´Ù.
HA Tag sequence: ACC ATG TAC CCA TAC GAT GTT CCA GAT TAC GCT
Myc Tag sequence: ATG GCA TCA ATG CAG AAG CTG ATC TCA GAG GAG GAC CTG
pCMV-Myc and pCMV-HA vector maps.