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

M084-5
Lot 012~Page 1

MONOCLONAL ANTIBODY

Anti-Podocalyxin (PCLP1) (Human) mAb-PE
Code No.
M084-5

Clone
53D11

Subclass
Mouse IgG2a

BACKGROUND: Recent studies with avian embryos

and murine embryonic stem cells have suggested that
hematopoietic cells are derived from hemangioblasts, thecommon precursors of hematopoietic and endothelial cells.
Hara et al. molecularly cloned podocalyxin-like protein 1(PCLP1) as a novel surface marker for endothelial-likecells in the AGM (aorta-gonad-mesonephros) region ofmouse embryos, where long-term repopulatinghematopoietic stem cells (LTR-HSCs) are known to arise.
PCLP1+CD45- cells in the AGM region incorporated
acetylated low-density lipoprotein and produced both
hematopoietic and endothelial cells when cocultured with
OP9 stromal cells. Moreover, multiple lineages ofhematopoietic cells were generated in vivo when PCLP1+
CD45- cells were injected into neonatal liver ofbusulfan-treated mice. Today it is reported that the PCLP1
is identical with the Podocalyxin.

SOURCE: This antibody was purified from mouse

Quantity
1 mL (50 tests)

SPECIES CROSS REACTIVITY:
Species

HumanMouse

Rat

Cell

HUVEC

Not tested

Not tested

Reactivity on FCM+REFERENCES:

1) Doyonnas, R., et al., Blood 105, 4170-4178 (2005)
2) Minegishi, N., et al., Blood 102, 896-905 (2003)
3) Schopperle, M.W., et al., Biochem. Biophys. Res. Commun.
300, 285-290 (2003)
4) Minehata, K., et al., Blood 99, 2360-2368 (2002)
5) Doyonnas, R., et al., J. Exp. Med. 194, 13-27 (2001)
6) Hara, T., et al., Immunity 11, 567-578 (1999)
7) Kershaw, B. D., et al., J. Biol. Chem. 272, 15708-15714 (1997)

RELATED PRODUCTS:

Please visit our website at https://ruo.mbl.co.jp/.

ascites fluid using protein A agarose. This hybridoma(53D11) was established by fusion of mouse myeloma cell
P3U1 with Balb/c mouse splenocyte immunized withCHOcell
expressing
full-length
human
Podocalyxin/PCLP1.

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 human
Podocalyxin/PCLP1 on Flow cytometry.

APPLICATION:

Flow cytometry; 20 µL (ready for use)
*Please refer to the data sheet (MBL, code no. M084-3)
for other applications.
Detailed

procedure

PROTOCOL.

INTENDED USE:

isprovidedinthefollowing

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

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

PROTOCOL:

Flow cytometric analysis for adherent cells

We usually use Fisher tubes or equivalents as reaction tubes
for all steps after 2).
1) Detach the cells from culture dish by using celldissociation buffer (Invitrogen, code no. 13151-014).
2) Wash the cells 3 times with washing buffer [PBS
containing 2% fetal calf serum (FCS) and 0.09% NaN3].
3) Resuspend the cells with washing buffer (5 x 106
cells/mL).

MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.

URL https://ruo.mbl.co.jp e-mail support@mbl.co.jp

M084-5Lot 012~Page 24) Add 50 µL of the cell suspension into each tube, andcentrifuge at 500 x g for 1 minute at room temperature.
Remove supernatant by careful aspiration.
5) Add 20 µL 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.
6) Add the primary antibody as suggested in theAPPLICATION. Mix well and incubate for 30 minutesat room temperature.
7) Add 1 mL of washing buffer followed by centrifugation
at 500 x g for 1 minute at room temperature. Remove
supernatant by careful aspiration.
8) Resuspend the cells with 500 µL of washing buffer
and analyze by a flow cytometer.
(Positive control for Flow cytometry; HUVEC)

240214-4