_SLIDE_1_
Protein concentration determination
_SLIDE_2_
´Ü¹éÁúÁ¤·®À̶õ
½Ã·á ³»¿¡ Æ÷ÇÔµÈ ´Ü¹éÁúÀÇ ¾ç, ¶Ç´Â ³óµµ¸¦ ±¸ÇÏ´Â °úÁ¤
_SLIDE_3_
¾ðÁ¦ ´Ü¹éÁú Á¤·®À» Çϴ°¡
wild type cell°ú mutant cell »çÀÌÀÇ Æ¯Á¤ protein¾ç ¹ßÇö¿¡ Â÷ÀÌ ¿©ºÎ¸¦ western blotÀ» ÀÌ¿ëÇؼ È®ÀÎÇÏ°í ½ÍÀ» °æ¿ì
¡æ total protein ¾çÀ» ´Ü¹éÁú Á¤·®À» ÀÌ¿ëÇؼ ¸ÂÃçÁØ´Ù.
why
¡æ total protein¾çÀÌ µ¿ÀÏÇؾß, ƯÁ¤ protein ¾çÀÇ »ó´ëÀûÀÎ °ªÀ» ºñ±³ÇÒ ¼ö Àֱ⠶§¹®ÀÌ´Ù.
2) enzyme activity¸¦ ÃøÁ¤ÇÒ °æ¿ì(enzyme-catalyzed reaction)
¡æ enzymeÀ» substrate¿¡ ó¸®ÇÑ ÈÄ, ´ÜÀ§ ½Ã°£´ç substrateÀÇ ³óµµ °¨¼Ò¸¦ ÅëÇÏ¿©, enzyme activity¸¦ ÃøÁ¤ÇÑ´Ù.
_SLIDE_4_
´Ü¹éÁúÀÇ ±¸Á¶
_SLIDE_5_
´Ü¹éÁú Á¤·® ¹æ¹ý
UV spectrophotometry
Biuret method
BCA method
Lowry method
Bradford method
_SLIDE_6_
UV spectrophotometry
¹æÇâÁ· ¾Æ¹Ì³ë»êÀÎ phe, tyr, trp¿¡ ÀÇÇÑ 280 nm
¿¡¼ ÃÖ´ë Èí±¤À» ÀÌ¿ëÇÑ ¹æ¹ý.
-50 ug - 1 mg/ml ¹üÀ§ÀÇ ´Ü¹éÁú¾çÀÇ ÃøÁ¤ÀÌ °¡´É
-ºü¸£°í ´Ü¹éÁú º¯¼ºÀÌ¡¦(»ý·«)
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