2. MATERIAL
Tip, Pippet, Test tube, spectro-20, D.W, ºñÄ¿, Centrifuge, Fecl3, H2SO4, Ethylacetate ethanol, »ï°¢Çø®½ºÅ©, Tritonx-100, cleanbanch
3. METHOD
1. Ç÷¾×À» »ó¿Â¿¡¼ ÀÀ°íµÉ¶§±îÁö ±â´Ù¸° ÈÄ ³ÃÀå°í¿¡¼ ³Ã°¢
2. ¾à 3000rpm 5min°¡ ¿ø½É ºÐ¸®
3. sup¸¸ µû·Î ¸ðÀº´Ù.
(1) cholesterol standard curve ÀÛ¼º
a.
Ç¥ÁØ¿ë¾×(1mg/ml)
0.5
1.0
1.5
2.0
2.5
ethylacetate
-etanol(ml)
4.5
4
3.5
3.0
2.5
½â¾î 3000rpm 5min ¿ø½ÉºÐ¸®
b. sup. 0.5ml + Fecl3 1ml + H2SO4 1mlÀ» ¼¯´Â´Ù.
c. A550¿¡¼ Èí±¤µµ ÃøÁ¤
d. standard curve ÀÛ¼º
(2) cholesterol ÃÑ·®
a. Ç÷û 0.5ml + ethylacetate ethanol 5ml¸¦ ¼¯´Â´Ù.
b. 3000rpm, 5ºÐ ¿ø½ÉºÐ¸®
c. sup. 0.5ml + Fecl3 1ml + H2SO4 1mlÀ» ¼¯´Â´Ù.
(3) free cholesterol ¾ç ÃøÁ¤
a. (2) ÃßÃâ¾× 0.6ml + Tritonx-100 2mlÀ» ¼¯´Â´Ù.
b. 10ºÐ°£ »ó¿Â¿¡¼ incubation
c¡¦(»ý·«)
e. acetone ¿ÏÀüÈ÷ Á¦°ÅÈÄ ethylacetate 0.3ml + Fecl3 1ml + H2SO4 1mlÀ» ¼¯´Â´Ù.
f. A550¿¡¼ Èí±¤µµ ÃøÁ¤
g. ¾ç ÃøÁ¤
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