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

D187-4
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MONOCLONAL ANTIBODY

Anti-Dlk (Pref-1) mAb-FITC
Code No.
D187-4

Clone
24-11

Subclass
Rat IgG1

BACKGROUND: Delta like protein (Dlk), also knownasPreadipocyte
factor-1(Pref-1)
orzonaglomerulosa-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 orinsulin 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.

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.

Quantity
100 L

Concentration
500 g/mL

SPECIES CROSS REACTIVITY:
Species

Human

Mouse

Rat

Cell

Not tested

Fetal
hepatocytes

Not tested

Reactivity on FCM+

REFERENCES:
1) Cheung, P. F., et al., PLoS One 6, e28246 (2011) [FCM]
2) Khurana, S. and Mukhopadhyay, A., Am. J. Pathol. 173,1818-1827 (2008) [FCM]
3) Suzuki, K., et al., Gastroenterology, 135, 270-281 (2008) [IHC]
4) Tanimizu, N., et al., J. Cell Sci. 116, 1775-1786 (2003)
5) Kaneta, M., et al., J. Immunol. 164, 256-264 (2000)
Clone 24-11 is used in the reference number 1)-3).

RELATED PRODUCTS:
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FORMULATION: 50 g IgG in 100 L volume of PBS
containing 1% BSA and 0.1% ProClin150.

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; 50 g/mL*Please refer to the data sheet (MBL, code no. D187-3)
for other applications.
Detailedprocedure

isprovided

inthefollowingPROTOCOL.
The descriptions of the following protocols are examples.
Each user should determine the appropriate condition.

INTENDED USE:
For Research Use Only. Not for use in diagnostic procedures.

PROTOCOL:
Flow cytometric analysis for floating cells
We usually use Fisher tubes or equivalents as reaction tubes
for all steps described below.

MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
URL https://ruo.mbl.co.jp

e-mail support@mbl.co.jp

D187-4
Lot 026~
Page 21) Wash the cells 3 times with washing buffer [PBScontaining 2% fetal calf serum (FCS) and 0.1% 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.

2) Resuspend the cells with washing buffer (5x10 6 cells/mL).
3) Add 50 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 Clear Back (human Fc receptor blocking
reagent, MBL, code no. MTG-001) to the cell pellet aftertapping. Mix well and incubate for 5 minutes at roomtemperature.
5) Add 40 L of the primary antibody at the concentration ofas suggested in the APPLICATION diluted in thewashing buffer. Mix well and incubate for 30 minutes atroom temperature.
6) Add 1 mL of the washing buffer followed bycentrifugation at 500 x g for 1 minute at room temperature.
Remove supernatant by careful aspiration.
7) Resuspend the cells with 500 L of the washing buffer
and analyze by a flow cytometer.
(Positive control for Flow cytometry; mouse fetal
hepatocytes, E14.5)

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