®

PlasmaSerum

1 to 2 1 to 4 1 to 8Plasma and Serum Dilutions

0.90.80.70.60.50.40.30.20.1

0

OD

630

nm

2.5

2

1.5

1

0.5

0

OD

540

nm

0 500 1000 1500 2000 2500 3000Lipid Standard (mg/dL)

0

200

400

600

800

1000

Relati

ve Rati

o

Exposure

Time

VAC01

02468

1012

Relative

Rati

o

Exposure Time

BAT10

NE(-) 1H 3H 6H 24H NE(-) 1H 3H 6H 24H

kDa

34

VAC01 VAC01+

NE6h

VAC01+

NE24h

BAT10

UCP1

43

GAPDH

26

2500

2000

1500

1000

500

0RF

U100 20 4 0

ヒト血清（％）

ヒト血清 (µL) 　　 ヒト血漿 (µL)50 12.5 3.125 0 50 12.5 3.125 0

2.42.2

21.81.61.41.2

10.80.60.40.2

0

OD

450

nm

Mouse Preadipocyte 3T3-L1Medium：D-MEM + 10%CS, D-MEM +10%FBS

Cell collection 2DG (1 mM) Insulin (1μM) Phloretin (200μM)

30min 48min 50min 70min

serumfree medium(D-MEM)

HKRP buffer

0-6hr

2DG 　 - + + +Insulin 　 - - + -Phloretin*1 　 - - - +

3000

2000

1000

2500

1500

500

0

2DG6

P in

corp

orat

ed in

to ce

lls (p

mol

e/w

ell)

*1 Phloretin : glucose 　 transporter inhibitor

serumfree medium(DE/F-12)

HKRP bufferWortmannin (50 nM) 2DG (1 mM) Insulin (1μM) Phloretin (200μM)

0 30min 48min 50min 70min

Cell collection(DE/F-12)

-6hr

Human Preadipocyte －subcutaneous fat tissue　　（ScienCell Research Laboratories Cat.# 7220）Medium ：Visceral Adipocyte Culture Medium ver.2 （Cat.# PMC-VACM2-COS）

2DG - + + + +Insulin - - + + +Phloretin - - - + -Wortmannin - - - - +

2DG6

P in

corp

orat

ed in

to ce

lls(p

mol

e/µg

DNA

)

800

600

400

200

0

*1 Phloretin : glucose transporter inhibitor　　*2 Wortmannin : PI3 kinase inhibitor

実験例 2　　ヒト皮下脂肪細胞による 2DGの取り込み

y = 407x + 4502.6R² = 1

0

5000

10000

15000

20000

25000

30000

0 10 20 30 40 50

Fluo

resc

ence

Inte

nsity

2DG6P(μM) 0

10

20

30

40

A B C

2DG

6P in

corp

orat

ed in

to c

ells

(nm

ole/

12 w

ell c

ultu

re d

ish)

添加物 Sample A Sample B Sample C

インスリン － － ＋

2DG － ＋ ＋

Denaturing extraction Native extraction

2509864503630

16

M MS.cer

evise

aeL.

Lacti

s (Gram

＋)

S.cer

evise

aefilam

entu

s

fungus

filamen

tus

fungus

250　　　350　　　450　　　550　　　650Wavelength (nm)

Excitaion Em

ission

Live-or-Dye 350/448 Live-or-Dye 405/452 Live-or-Dye 405/545 Live-or-Dye 488/515

Live-or-Dye 568/583 Live-or-Dye 594/614 Live-or-Dye 640/662 Live-or-Dye 750/777

Live-or-Dye 488/515 Live-or-Dye 568/583

Live-or-Dye 594/614 Live-or-Dye 640/662

0

50

100

150

0 5 10 15Albu

min

pro

duct

ion

(µg/

day/

wel

l)

Culture time (day)

Plastic Plate/5%-O2

VECELL® G-Plate/5%-O2

Plastic Plate/21%-O2

VECELL® G-Plate/21%-O2

Plastic Plate/5→21%-O2

VECELL® G-Plate/5→21%-O2

3

15

12

9

6

3

06 10

Reso

rufin

(pm

ol/h

/wel

l)

Culture time (Day)

Plastic Plate/O2-

VECELL® H-Plate/O2+

ScienCellResearch Laboratories

TM

®

1

2

3

4

5 6 7

サンプル希釈液マウス血清マウス血漿組織培養培地他社製品

マウス IFN-β (pg/㎖)0.1 1 10 100 1000

吸光

度 【

45

0nm

】

3

2

1

0

pg/㎖

OD

45

0nm

5

0.5

0.050.5 5 50

Mammalian Mu Beta

E.coli Mu Beta