First, the organization sample
1
Also use isotonic buffer, by weight
1
:
9
, preparing a homogenate,
2500rpm*10min
, take the supernatant
10%
Homogenate
TG
,
TC
Determination (extraction is not complete, but can also be measured)
2
Extracted with an organic solvent,
a:
Using anhydrous ethanol as the homogenization medium, the homogenate was prepared according to the above method, and the extraction effect was better than the isotonic buffer.
b:
Extraction with special organic solvents (such as methanol mixture, etc.) is good, but due to high volatilization, the measurement is not accurate, so it is generally required.
N2
After drying, it was reconstituted with absolute ethanol and then measured.
Second, culture cells
1
,use
PBS
Washing cells
2
Join
500ul
Ethanol, so that it is spread over the culture dish. When the cells in the culture dish are white, the culture dish is placed obliquely, and the cells are scraped off with a cell scraper to soak the cells in ethanol.
3
Transfer the cells and ethanol to the glass homogenizer with a pipette and grind
3
Minute into suspension
4
, collecting the suspension,
4000rpm*10min
Centrifuge, take the supernatant
50ul
Determination
Third, the milk sample
Fresh or frozen milk samples are extracted with absolute ethanol in different proportions (milk sample: absolute ethanol)
=1
:
1
,
1
:
4
,
1
:
9
,
1
:
19
,
1
:
49
,
1
:
99
Full vortex to mix
2
minute,
4000
turn
/
Centrifugal
10
Minutes, take the clearing and press the operation table.
According to the dilution and the result, the concentration curve is taken, and the concentration of the linear linear segment is suitable.
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