For Research Use Only.
Not for use in diagnostic procedures.

D187-5
Page 1 of 2

MONOCLONAL ANTIBODY

Anti-Dlk (Pref-1) mAb-PE
Code No.
D187-5

Clone
24-11

BACKGROUND: Delta like protein (Dlk), also knownasPreadipocyte
factor-1(Pref-1)or
zona
glomerulosa-specific factor (ZOG), is an EGF-like
transmembrane protein expressed preadipocytes but not inmature adipocytes. It is highly expressed in fetal liver, theadrenal gland, and placenta, as well as someneuroendocrine tumors and small cell lung carcinomas,
where it plays a role in differentiation and proliferation.
Dlk positively and negatively regulates adipocyte
differentiation via at least four major variants (45-60 kDa)
of Dlk generated by alternatively splicing. Constitutive
expression of Dlk inhibits adipogenesis, but insulin or
insulin like growth factor-1 (IGF-1) can circumvent thisinhibition. Regulated processing of Dlk releases a 50 kDa
soluble form that was previously characterized as Fetal
Antigen-1, a protein involved in pancreatic island celldifferentiation.

SubclassRat IgG1

Quantity
1 mL (50 tests)

SPECIES CROSS REACTIVITY:
Species

HumanMouse

RatCell

Not tested

Fetalhepatocytes

Not tested

Reactivity on FCM+INTENDED USE:
For Research Use Only. Not for use in diagnostic procedures.

SOURCE: This antibody was purified from hybridoma(clone 24-11) supernatant using protein G agarose. Thishybridoma was established by fusion of mouse myeloma
cell with rat splenocyte immunized with Pref-1-Fc protein.

FORMULATION: 50 tests in 1 mL volume of PBS
containing 1% BSA and 0.09% NaN3.
*Azide may react with copper or lead in plumbing system toform explosive metal azides. Therefore, always flush plenty ofwater when disposing materials containing azide into drain.

STORAGE: This antibody solution is stable for one year
from the date of purchase when stored at 4°C.

REACTIVITY: This antibody reacts with Dlk on Flowcytometry.

APPLICATION:

Flow cytometry; 20 L (ready for use)
*Please refer to the data sheet (MBL code no. D187-3)
for other applications.
Detailed

procedure
PROTOCOL.

isprovided

inthefollowing

REFERENCES:
1) Takeshita, S., et al., J. Biol. Chem. 289, 16699-16710 (2014)
2) Tanimizu, N., et al., J. Cell Sci. 116, 1775-1786 (2003)
3) Kaneta, M., et al., J. Immunol. 164, 256-264 (2000)

Flow cytometric analysis of Dlk expression
on mouse Fetal Hepatocytes. Openhistogram indicates the reaction of isotypic
control to the cells. Shaded histogramindicates the reaction of D187-5 to the cells.
PROTOCOL:
Flow cytometric analysis for floating cells
We usually use Fisher tubes or equivalents as reaction tubes
for all steps described below.
1) Wash the cells 3 times with washing buffer [PBS
containing 2% fetal calf serum (FCS) and 0.09% NaN3].
2) Resuspend the cells with washing buffer (1 x 107cells/mL).
3) Add 100 L of the cell suspension into each tube, andcentrifuge at 500 x g for 1 minute at room temperature(20~25°C). Remove supernatant by careful aspiration.
4) Add 10 L of normal goat serum containing 0.1% NaN3
to the cell pellet after tapping. Mix well and incubate for5 minutes at room temperature
5) Add the primary antibody as suggested in theAPPLICATION to the each tube. Mix well and incubatefor 15 minutes at room temperature.
6) Add 1 mL of the washing buffer followed bycentrifugation at 500 x g for 1 minute at roomtemperature. Remove supernatant by careful aspiration.
7) Resuspend the cells with 500 L of the washing buffer
and analyze by a flow cytometer.

MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
URL http://ruo.mbl.co.jp
e-mail support@mbl.co.jp, TEL 052-238-1904

D187-5
Page 2 of 2

(Positive control for Flow cytometry; Mouse fetal
hepatocytes, E14.5)

RELATED PRODUCTS:
D187-3
D187-4
M080-3
M080-4
M080-5Anti-Dlk (Pref-1) mAb (24-11)
Anti-Dlk (Pref-1) mAb-FITC (24-11)
Rat IgG1 (isotype control) (1H5)
Rat IgG1 (isotype control)-FITC (1H5)
Rat IgG1 (isotype control)-PE (1H5)150930-2