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˜µ¦µŠš¸É 1 °µ®µ¦š—¨°Š
„µ¦´œš¹„ o°¤¼¨
´œš¹„œÊ宜´„˜´ ª °ŠÅ„nÁœºÊ°ÄœÂ˜n¨³ŽÊεĜª´œš¸É 0, 4, 7, 14 ¨³ 21 °Š„µ¦š—¨°Š ´œš¹„ž¦·¤µ–°µ®µ¦
š¸É„·œÄœÂ˜n¨³ª´œ Á¡ºÉ°«¹„¬µ¤¦¦™£µ¡„µ¦Á‹¦·Á˜·Ã˜Â¨³ž¦³·š›·£µ¡„µ¦Äo°µ®µ¦
„µ¦Á˜¦¸¥¤˜´ª°¥nµŠ
Ĝª´œš¸É 4, 7, 14 ¨³ 21 ª´œ šÎ µ„µ¦Á„ȝ˜´ª°¥nµŠ˜´°n°œ °ŠÅ„n ÁœºÊ°ŽÊ娳 4 ˜´ª ®¨´Š‹µ„œ´ÊœÁ˜·¤ 50mM Tris
– HCl buffer pH 8.0 Ĝ˜´ª°¥n µŠ˜´°n°œ Ĝ°´˜¦µnªœ °Š˜´ª°¥nµŠ˜n °´¢Á¢°¦r 1:2 (w/v) ¨oª—˜´ª°¥nµŠÄ®o¨³Á°¸ ¥—
ץčo homogenizer ‹µ„œ´ÊœœÎµ˜´ª°¥nµŠš¸É—Å—oޞ{~œ—oª¥Á‡¦ºÉ°ŠÁ®ª¸É ¥ŠÂ¦Š¼Šš¸É‡ªµ¤Á¦È ª 10000 g š¸É°»–®£¼ ¤·
4 °Š«µÁŽ¨ÁŽ¸¥œµœ 15 œµš¸ Â¥„Á°µÁŒ¡µ³nªœ¨°¥ (supernatant) Á„ȝŪoš¸É – 20 °Š«µÁŽ¨ÁŽ¸¥ (Gimenez et al.,
1999) Á¡ºÉ°Äo Ĝ„µ¦ª·Á‡¦µ³®r „·‹„¦¦¤ °ŠÁ°œÅŽ¤rŨÁž
„µ¦«¹„¬µ„·‹„¦¦¤ °ŠÁ°œÅŽ¤r ŨÁž
„·‹„¦¦¤ °ŠÁ°œÅŽ¤r ŨÁž«¹„¬µÃ—¥Äoª·›¸„µ¦ °Š Markweg – Hank (1995) čo 0.01 M para–
nitrophenylpalmitate (pNNP) 100 ôl ˜´ª°¥n µŠÁ°œÅŽ¤rŨÁžš¸É „´—‹µ„˜´°n°œ 50 ôl ¨³ 50mM Tris – HCl buffer
800 ôl Ÿ¤Ä®oÁ oµ„´œ—¸Â¨oªœÎµÅžn¤š¸É°»–®£¼¤· 37 °Š«µÁŽ¨ÁŽ¸¥œµœ 60 œµš¸ ‹µ„œ´ÊœÁ˜· ¤ 0.1M Na2CO3 250 ôl
¨oªœÎ µÅžž{~œ—o ª¥Á‡¦ºÉ°ŠÁ®ª¸É¥ŠÂ¦Š¼Šš¸É‡ªµ¤Á¦Èª 10000 g š¸É°»–®£¼¤· 4 °Š«µÁŽ¨ÁŽ¸ ¥œµœ 15 œµš¸ œÎ µÅžª´—‡nµ„µ¦
—¼—„¨ºœÂŠš¸É‡ªµ¤¥µª‡¨ºÉœÂŠ 410 nm —o ª¥Á‡¦ºÉ°Š spectrophotometer
„µ¦ª· Á‡¦µ³®rž¦·¤µ–Þ¦˜¸œÄoª·›¸ Lowry’s method (1915) čo bovine serum albumin Áž}œµ¦Ãž¦˜¸œ
¤µ˜¦“µœ
„¨»n¤š—¨°Š °µ®µ¦š—¨°Š
1. ‡ª‡»¤ (T1) °µ®µ¦¡ºÊœ“µœ
2. Å ¤´œ¼Š (T2) °µ®µ¦Å ¤´œ¼Š
3. Á¨Ž·˜·œ (T3) °µ®µ¦Å ¤´œ¼Š + Á¨Ž·˜·œ 0.50%
4. ŸŠœÊε—¸ »„¦ 1 (T4) °µ®µ¦Å ¤´œ¼Š + ŸŠœÊÎ µ—¸ »„¦ 0.125%
5. ŸŠœÊε—¸ »„¦ 2 (T5) °µ®µ¦Å ¤´œ¼Š + ŸŠœÊÎ µ—¸ »„¦ 0.25%
6. ŸŠœÊε—¸ »„¦ 3 (T6) °µ®µ¦Å ¤´œ¼Š + ŸŠœÊÎ µ—¸ »„¦ 0.50%