K0224-A48 For Research Use Only. Page 1 of 2 Not for use in diagnostic procedures.

MONOCLONAL ANTIBODY Alexa Fluor® 488 labeled Anti-Human -3/CD113

Code No. Clone Subclass Quantity Concentration K0224-A48 N3.12.4 Mouse IgG2a  100 g 1 mg/mL

BACKGROUND: originally identified as REFERENCE: poliovirus receptor-related (PRR) are 1) Takai, Y., and Nakanishi, H., J. Sci. 116, 17-27 (2003) Ca2+-independent immunoglobulin-like cell-cell-adhesion molecules. Nectins comprise a family of four members: nectin-1 (CD111/PRR1), nectin-2 (CD112/PRR2), nectin-3 (CD113/PRR3) and nectin-4 (PRR4). Nectin-2 and -3 are ubiquitously expressed whereas nectin-1 is abundantly expressed in brain, and nectin-4 is in placenta. Nectins colocalized with afadin, which an -filament binding protein to associated with the actin . Nectins play important roles in diverse cell-cell junctions.

SOURCE: This antibody was purified from hybridoma (clone N3.12.4) supernatant using protein A agarose. The mouse was immunized with recombinant human Nectin-3 Flow cytometric analysis of human Fc fusion protein. Nectin-3 expression on U251. Open histogram indicates the reaction of isotypic control to the cells. Shaded histogram FORMULATION: 100 g IgG in 100 L volume of indicates the reaction of K0224-A48 to the PBS containing 1% BSA and 0.09% NaN3. cells. *Azide may react with copper or lead in plumbing system to form explosive metal azides. Therefore, always flush plenty of water when disposing materials containing azide into drain. PROTOCOL: Flow cytometric analysis for floating cells STORAGE: This antibody is stable for one year from the We usually use Fisher tubes or equivalents as reaction tubes o date of shipment when stored at 4 C. for all steps described below. 1) Wash the cells 3 times with washing buffer [PBS REACTIVITY: This antibody reacts with V domain of containing 2% fetal calf serum (FCS) and 0.1% NaN3]. 6 human Nectin-3 on Flow cytometry. 2) Resuspend the cells with washing buffer (5x10 cells/mL). 3) Add 50 L of the cell suspension into each tube, and APPLICATIONS: centrifuge at 500 x g for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration. Flow cytometry; 10 g/mL (final concentration) 4) Add 20 L of Clear Back (human Fc receptor blocking Immunocytochemistry; Not recommended reagent, MBL; code no. MTG-001) to the cell pellet after *Please refer to the data sheet (MBL code no. K0224-3) tapping. Mix well and incubate for 5 minutes at room for other applications. temperature.

5) Add 40 L of the primary antibody at the concentration as Detailed procedure is provided in the following suggested in the APPLICATIONS diluted in the washing PROTOCOL. buffer. Mix well and incubate for 30 minutes at room temperature. SPECIES CROSS REACTIVITY: 6) Add 1 mL of the washing buffer followed by Species Human Mouse Rat centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration. Cell U251 Not Tested Not Tested 7) Resuspend the cells with 500 L of the washing buffer and analyze by a flow cytometer. Reactivity on FCM + (Positive control for Flow cytometry; U251)

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

MEDICAL & BIOLOGICAL LABORATORIES CO., LTD. URL https://ruo.mbl.co.jp e-mail [email protected], TEL 052-238-1904 K0224-A48 Pa ge 2 of 2

RELATED PRODUCTS: K0224-3 anti-human Nectin-3/CD113 (N3.12.4) D175-3 CD112/Nectin-2 (TX31) D175-4 FITC labeled CD112/Nectin-2 (TX31) D146-3 anti-mouse Nectin-1 (48-12) D083-3 Mouse CD112/Nectin-2 (502-57) D084-3 Mouse CD113/Nectin-3 (103-A1) M076-A48 Alexa Fluor® 488 labeled Mouse IgG2a isotype control (6H3)

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LABEL LICENSES: This product is provided under an agreement between TECHNOLOGIES Corporation, and Medical & Biological Laboratories Co., LTD, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications, and corresponding non-U.S. equivalents, owned by or licensed to LIFE TECHNOLOGIES Corporation. For information on purchasing a license to this product for any other use, contact LIFE TECHNOLOGIES Corporation, Molecular Probes Detection Technologies, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc.