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zhuzhaofu

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alpha - In the grape glycosidase suppression active determination 100 μL reacting system the right amount mammal originates the grape glycosidase (α-glucosidase-rat, withdraws, 67 nM sodium phosphate buffer solution by big mouse small intestine organization in) (pH 6.8) and the sample, simultaneously sets up the blank comparison (not including enzyme and sample) and the negative comparison (not including sample), 37℃ responds 10min, joins 0.1 M malt sugar, the room temperature responded 10 min, joins 200 μL glucose examination reagent again, mixes uniform latter 490 nM to determine the OD value.According to OD value computation suppression rate, suppression rate = [1- (OD sample - OD blank)/(OD negative - OD blank)]¡Á100.Each sample each density supposes the double duplicate hole, duplicates two times.
PPAR responded the part (PPRE) excited activeness examination experiments the HepG2 liver cancer cell convention raise in 37℃, 5%CO2, contains 100U•mL-1 chain mildew element and in penicillin low sugar DMEM.1.5¡Á104 a/vaccination raises the over night after 96 orifices, the reference extension dyes the reagent instruction booklet to carry on the material particle extension to dye.The extension dyes the material particle including has the PPAR response part and firefly fluorescein enzyme report gene material particle pPPRE-Luc, and serves as the extension to dye the materials for internal reference to illuminate has sea kidney fluorescein enzyme material particle phRL-TK, after the extension dyes 24h to use in exchange including treats measured (cell which the sample the culture medium, simultaneously sets up (which the blank comparison not to have transferred dyes cell), the negative comparison extension dyes not to add (cell which sample) and masculine comparison extension dyes to join the Geleg alkone, Pioglitazone).After continues to raise 24h with the double fluorescein enzyme report gene examination reagent box (Promega) examination fluorescein enzyme activity, according to chemiluminescence intensity L value computation which examines excited rate.L1 is the firefly fluorescein enzyme chemiluminescence intensity, L2 is the materials for internal reference according to the sea kidney fluorescein enzyme chemiluminescence intensity, excited rate = [(L1 sample - L1 blank)/(L1 negative - L1 blank)]/[(L2 sample - L2 blank)/(L2 negative - L2 blank)]¡Á100%; Sample examination density for 10μg•mL-1, when examination supposes the double duplicate hole, duplicates two times.
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goodtimega

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linghanyuan(½ð±Ò+2,VIP+0):лл½»Á÷ 9-27 07:35
¦Á-glucosidase inhibitory activity is determined through 100 ¦ÌL reaction system suitable mammalian source glucosidase (¦Á-glucosidase-rat that was extracted from the rat small intestine)with 67 nM sodium phosphate buffer (pH 6.8). This sets up a blank control (without enzyme and samples) and negative control (without sample) through 37 ¡æ reaction to 10min, adding 0.1 M maltose, after room temperature reaction of 10 min, then adding 200 ¦ÌL of glucose testing reagents, mixing post-490 nM measured OD values. Inhibition rate calculated is in accordance to OD values of the inhibition rate = [1 - (OD sample-OD blank) / (OD negative-OD blank)] ¡Á 100. The concentration of each sample shows a double recovery for each hole after being repeated twice.

PPAR response element (PPRE) activity in emotion-testing of conventional HepG2 hepatoma cells was cultured in 37 ¡æ, 5% CO2, containing 100U • mL-1 streptomycin and penicillin in the sugar DMEM. 1.5 ¡Á 104 / holes were inoculated into 96-well plates cultured overnight, taking into account the instructions for plasmid transfection reagent transfection. A plasmid was transferred including the PPAR-responsive element and the firefly luciferase reporter gene plasmid pPPRE-Luc, and being used as a reference within transfected with a Renilla luciferase plasmid phRL-TK, and transfection 24h.  After changing the medium containing the sample under test, the establishment of blank control (non-transfected cells), negative control (cells transfected without samples), and positive control (cells transfected by adding Pioglitazone, Pioglitazone) was achived. This process was continued to foster 24h later Dual-Luciferase Reporter Gene Assay Kit (Promega) through detection of luciferase activity, according to detected values of chemiluminescence intensity of L calculated the rate of excited. L1 stands for the firefly luciferase chemiluminescence intensity, L2 as an internal reference Renilla luciferase chemiluminescence intensity, excitement rate = [(L1 Sample-L1 blank) / (L1-negative-L1 blank )]/[( L2 Sample -L2 blank) / (L2-negative-L2 blank)] ¡Á 100%.  Samples detected concentrations of 10¦Ìg • mL-1, a double re-hole detection were repeated twice.
2Â¥2009-09-27 02:18:48
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yxzhang838

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Originally posted by goodtimega at 2009-9-27 02:18:
¦Á-glucosidase inhibitory activity is determined through 100 ¦ÌL reaction system suitable mammalian source glucosidase (¦Á-glucosidase-rat that was extracted from the rat small intestine)with 67 nM ...

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4Â¥2009-09-27 19:33:15
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goodtimega

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Originally posted by yxzhang838 at 2009-9-27 19:33:

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