Monoclonal Antibody to RANK (CD265/ODFR

SM7086P OriGene Technologies Inc. OriGene EU

Acris Antibodies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-888-267-4436 Phone: +49-5221-34606-0 Fax: +1-301-340-8606 Fax: +49-5221-34606-11 [email protected] [email protected]

Monoclonal Antibody to RANK (CD265/ODFR) - Purified Alternate names: ODFR, Osteoclast differentiation factor receptor, Receptor activator of NF-KB, TNFRSF11A, Tumor necrosis factor receptor superfamily member 11A Catalog No.: SM7086P Quantity: 0.1 mg Concentration: 0.5 mg/ml Background: Apoptosis, or programmed cell death, occurs during normal cellular differentiation and development of multicellular organisms. Apoptosis is induced by certain cytokines including TNF and Fas ligand of the TNF family through their death domain containing receptors, TNFR1 and Fas. Receptor activator of NF-kB (RANK) is a recently cloned member of the TNFR superfamily with no significant homology to other members of this family (1). RANK ligand (RANKL/TRANCE/ OPGL) binds to RANK on dendritic cells, upregulates the expression of anti-apoptotic protein BcL-XL suggesting a role in dendritic cell survival (2). The cytoplasmic domain of RANK interacts with TRAF2, TRAF5 and TRAF6 (3,4). Overexpression of RANK activates NF-kB and c-Jun-terminal kinase (JNK) pathways (3). Recent studies have shown that RANK interaction with TRAF6 activates NF-kB, whereas JNK activation is mediated through binding of RANK to TRAF2 (4). Uniprot ID: Q9Y6Q6 NCBI: NP_003830.1 GeneID: 8792 Host / Isotype: Mouse / IgG1 Recommended SM10P (for use in human samples), AM03095PU-N Isotype Controls: Clone: 9A725 Immunogen: Fusion protein containing Amino Acid residues 326-616 of human RANK. Format: State: Liquid purified IgG fraction Purification: Protein G Chromatography Buffer System: PBS Preservatives: 0.05% Sodium Azide Stabilizers: 0.05% BSA Applications: Flow Cytometry (Intracellular): 2-4 µg/ml. Immunohistochemistry on Paraffin Sections (Ref.5, see also Protocol below). Western Blot: 1-2 µg/ml. Raw 264 Cells can be used as Positive Control, detects a protein of approx. 97 kDa. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

For research and in vitro use only. Not for diagnostic or therapeutic work. MS/20140910 Material Safety Datasheets are available at www.acris-antibodies.com or on request. Acris Antibodies is now part of the OriGene family. Learn more at www.origene.com 1 / 2 SM7086P: Monoclonal Antibody to RANK (CD265/ODFR) - Purified Specificity: This antibody recognizes an epitope located between amino acid residues 330-427 of Human RANK. Species: Human and Mouse. Other species not tested. Storage: Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: One year from despatch. General Readings: 1. Anderson et al. Nature 390: 175-179 (1997). 2. Wong, B.R et al. J. Exp. Med. 186: 2075-2080. 3. Darnay, B.G et al. J. Biol. Chem 273: 20551-20555 (1998). 4. Darnay, B.G et al. J. Biol. Chem. 274: 7724-7731 (1999). 5. Fiumara, P., Snell, V., Li, Y., Mukhopadhyay, A., Younes, M., Gillenwater, A., Cabanillas, F., Aggarwal, B., Younes, A. Blood, 98: 2784-2790 (2001). 6. Bharti AC et al. J. Biol. Chem., Vol. 279, Issue 7, 6065-6076 (2004). Protocols: Protocol for Paraffin Sections: 1. Prepare paraffin-embedded sections. 2. Immunostain sections using a Techmate 1000 automatic immunostainer (Ventana, Tucson, AZ). Deparaffinize sections in xylene, rehydrate with decreasing concentrations of alcohol and finally immerse in phosphate-buffered saline (PBS). 3. Perform antigen retrieval using steam-heat in 10 mM citrate buffer (pH 6.0) for 30 minutes in a commercially available vegetable steamer. 4. Rinse sections in distilled water, wash in PBS for 5 minutes, and incubate for 15 minutes with DAKO protein block (DAKO, Carpenteria, CA). 5. Incubate sections for 2 hours with SM7086P, diluted 1:500 in 0.1% bovine serum albumin/PBS. 6. Wash sections and detect using an LSAB2 peroxidase kit (primary rabbit/mouse; DAKO) with diaminobenzidine as the chromogen. 7. Counterstain sections with hematoxylin. 8. Dehydrate and mount. Pictures: Figure 1. Western blot analysis of RANK in 293 cells transfected with RANK cDNA. A single protein band with an approximate Mol. wt. of 97 kDa is detected.

Flow Analysis in RAW cells using 2ug of RANK antibody. Shaded histogram represents RAW cells without antibody; green represents isotype control; purple represents anti-RANK antibody.