BD Pharmingen™

Technical Data Sheet

Purified Mouse Anti-Human Disialoganglioside GD3Product Information

554274Material Number:

Ganglioside GD3Alternate Name:

Size: 0.1 mg

Concentration: 0.5 mg/ml

Clone: MB3.6

Immunogen: FM9 human melamona cell line

Isotype: Mouse IgG3, κ

QC Testing: HumanReactivity:

Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium

azide.

DescriptionGangliosides are sialic-acid bearing glycolipids expressed on the surface of all mammalian cells, and are likely involved in mediating

cell-substratum interactions. They are important target antigens for antibody-mediated cytolysis of human melanoma and neuroblastoma cells.

Human melanoma cells produce gangliosides GD2 and/or GD3 which are present in substratum-attached material, and may play a significant

role in the melanoma metastatic phenotype. Ganglioside GD3 is a major surface marker on most human melanoma cells. MB3.6 has been used

to localize GD3 in the plasma membrane and in focal adhesion plaques of human melanoma cells. Clone MB3.6 has also been shown to lyse

GD3 positive human melanoma cells by both antibody-dependent and complement-mediated cytotoxicity, as well as inhibit the growth of

human melanoma cells in athymic nude mice. Clone MB3.6 specifically reacts with human GD3 ganglioside. It does not cross-react with a

variety of other gangliosides purified from melanoma or neuroblastoma cells. The FM9 human melanoma cell line was used as the

immunogen.

Flow cytometric analysis of GD3 expression on M21

human melanoma cells. Melanoma cells were stained with

either Purified Mouse Anti-Human Disialoganglioside GD3

(Cat. No. 554274; filled histogram), or with Purified Mouse

IgG3, κ Isotype Control (Cat. No.553486; empty histogram).

Secondary staining was carried out with FITC Goat

Anti-Mouse IgG/IgM (Cat. No. 555988).The fluorescence

histograms were derived from gated events with the forward

and side light-scattering characteristics of viable melanoma

cells. Flow cytometry was performed on a BD FACScanTM.

Preparation and StorageStore undiluted at 4°C.

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Application Notes

Application

Flow cytometry Routinely Tested

Immunohistochemistry-frozen Reported

Immunofluorescence Reported

Cytotoxicity Reported

Page 1 of 2 554274 Rev. 2

Recommended Assay Procedure:

Applications include flow cytometry (1-2 µg/1x10^6 cells). Other applications not routinely tested at BD Biosciences Pharmingen include

immunohistochemistry of frozen tissue sections, immunofluorescence microscopy of cultured cells and antibody-dependent and complement

mediated cytotoxicity of GD3 positive cells.

Suggested Companion Products

Catalog Number Name CloneSize

555988 FITC Goat Anti-Mouse IgG/IgM 0.5 mg Polyclonal

553486 Purified Mouse IgG3, κ Isotype Control 0.5 mg A112-3

554656 Stain Buffer (FBS) 500 mL (none)

554657 Stain Buffer (BSA) 500 mL (none)

Product NoticesSince applications vary, each investigator should titrate the reagent to obtain optimal results. 1.

An isotype control should be used at the same concentration as the antibody of interest. 2.

Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before

discarding to avoid accumulation of potentially explosive deposits in plumbing.

3.

Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not

be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or

dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the

NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.

4.

Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols. 5.

ReferencesCheresh DA, Harper JR, Schulz G, Reisfeld RA. Localization of the gangliosides GD2 and GD3 in adhesion plaques and on the surface of human melanoma cells.

Proc Natl Acad Sci U S A. 1984; 81(18):5767-5771. (Immunogen: Cytotoxicity, Immunofluorescence)

Cheresh DA, Honsik CJ, Staffileno LK, Jung G, Reisfeld RA. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal

antibody-mediated tumor cytolysis. Proc Natl Acad Sci U S A. 1985; 82(15):5155-5159. (Clone-specific: Flow cytometry)

Cheresh DA, Klier FG. Disialoganglioside GD2 distributes preferentially into substrate-associated microprocesses on human melanoma cells during their

attachment to fibronectin. J Cell Biol. 1986; 102(5):1887-1897. (Biology)

Cheresh DA, Pierschbacher MD, Herzig MA, Mujoo K. Disialogangliosides GD2 and GD3 are involved in the attachment of human melanoma and neuroblastoma

cells to extracellular matrix proteins. J Cell Biol. 1986; 102(3):688-696. (Biology)

Hakomori S. Tumor-associated carbohydrate antigens. Annu Rev Immunol. 1984; 2:103-126. (Biology)

Mujoo K, Cheresh DA, Yang HM, Reisfeld RA. Disialoganglioside GD2 on human neuroblastoma cells: target antigen for monoclonal antibody-mediated cytolysis

and suppression of tumor growth. Cancer Res. 1987; 47(4):1098-1104. (Biology)

Page 2 of 2 554274 Rev. 2