ANTI-DC-STAMP CLONE 1A2

General description

Dendritic cell-specific transmembrane protein (UniProt Q9H295; also known as DC-STAMP, Dendrocyte-expressed seven transmembrane protein, FIND, hDC-STAMP, IL-four-induced protein, Transmembrane 7 superfamily member 4) is encoded by the DCSTAMP (also known as TM7SF4) gene (Gene ID 81501) in human. DC-STAMP is a six-transmembrane protein essential for cell-to-cell fusion to form multinucleated osteoclasts (OCs) during osteoclastogenesis. DC-STAMP expression is upregulated among osteoclast precursor (OCP) cells upon exposure to OC-promoting cytokines, such as receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL), and Dcstamp-knockout (KO) mice have few multinucleated TRAP+ OCs and increased bone mass. On the other hand, DC-STAMP overexpression in transgenic (Tg) mice causes accelerated cell-to-cell fusion during OCP differentiation and enhanced bone resorption. DC-STAMP is a six-trasmembrane (a.a. 35-55, 58-78. 98-118, 210-230, 293-313, 377-397) protein, having both its N- and C-terminal ends exposed intracellularly (a.a. 1-34, 398-470). The C-terminal cytoplasmic tail of DC-STAMP contains an immunoreceptor tyrosine-based inhibitory motif or ITIM sequence (407-SFYPSV-412) that, when phosphorylated on the tyrosine residue, recruits SHP-1. DC-STAMP neutralizing antibody blocks OC formation in vitro and abolishes cellular DC-STAMP and SHP-1 tyrosine phosphorylation.

Specificity

Clone 1A2 recognizes an epitope conserved between human and murine DC-STAMP in the third extracellular domain.

Immunogen

Epitope: The third extracellular domain.

KLH-conjugated linear peptide corresponding a sequence from the third extracellular domain of human DC-STAMP.

Application

Anti-DC-STAMP Antibody, clone 1A2 is an antibody against DC-STAMP for use in Western Blotting, Flow Cytometry, Immunocytochemistry, Immunoprecipitation.

Research Category
Inflammation & Immunology

Research Sub Category
Osteobiology

Western Blotting Analysis: 4.0 µg/mL from a representative lot detected DC-STAMP in 10 µg of thymus lysates from wild-type, but not Dcstamp-knockout mice.
Western Blotting Analysis: 1.0 µg/mL from a representative lot detected DC-STAMP in thymus and spleen lysates from wild-type mice, and greatly reduced DC-STAMP in thymus and spleen lysates from Dcstamp-knockout mice (Courtesy of Grace Chiu, Ph.D., University of Rochester Medical Center, NY, USA).
Western Blotting Analysis: A representative lot detected the ~106 kDa dimeric and the ~53 kDa monomeric DC-STAMP band by Western blotting under non-denatured and denatured condition, respectively, following DC-STAMP immunoprecipitation using murine RAW 264.7 macrophage lysate (Mensah, K.A., et al. (2010). J. Cell. Physiol. 223(1):76-83).
Flow Cytometry Analysis: A representative lot detected DC-STAMP-positive lymphoctes and monocytes in purified human PBMCs (Chiu, Y.H., et al. (2012). J. Bone Miner. Res. 27(1):79-92).
Flow Cytometry Analysis: A representative lot detected DC-STAMP surface expression on murine RAW 264.7 macrophages and murine bone marrow-derived CD11b+ monocytes. DC-STAMP is expressed on osteoclast precursor (OCP) cells as a dimer, which is efficiently detected by flow cytometry using clone 1A2 (Mensah, K.A., et al. (2010). J. Cell. Physiol. 223(1):76-83).
Function Analysis: A representative lot inhibited RANKL & M-CSF treatment-induced osteoclasts (OC) formation in human PBMCs & monocytes cultures (Chiu, Y.H., et al. (2012). J. Bone Miner. Res. 27(1):79-92).
Function Analysis: A representative lot inhibited RANKL treatment-induced osteoclasts (OC) formation in RAW 264.7 and murine bone marrow macrophages cultures (Mensah, K.A., et al. (2010). J. Cell. Physiol. 223(1):76-83).
Immunohistochemistry Analysis: A representative lot detected DC-STAMP expressionon in multinucleated ‘osteoclast-like’ giant cells in human giant cell tumor of bone (Chiu, Y.H., et al. (2012). J. Bone Miner. Res. 27(1):79-92).
Immunocytochemistry Analysis: A representative lot detected DC-STAMP-positive human PBMCs using 10% NBF-fixed, paraffin-embedded cell preparation (Chiu, Y.H., et al. (2012). J. Bone Miner. Res. 27(1):79-92).
Immunocytochemistry Analysis: A representative lot detected differential DC-STAMP intracellular localizations by fluorescent immunocytochemistry staining of 4% paraformaldehyde-fixed, 0.1% saponin-permeabilized murine bone marrow macrophages at different time points during osteoclastogenesis upon RANKL treatment (Mensah, K.A., et al. (2010). J. Cell. Physiol. 223(1):76-83).
Immunoprecipitation Analysis: A representative lot immunoprecipitated DC-STAMP from the lysates of human monocytes (Chiu, Y.H., et al. (2012). J. Bone Miner. Res. 27(1):79-92).
Immunoprecipitation Analysis: A representative lot immunoprecipitated DC-STAMP from the membrane extracts of murine RAW 264.7 macrophages (Mensah, K.A., et al. (2010). J. Cell. Physiol. 223(1):76-83).

Quality

Evaluated by Western Blotting in mouse thymus lysate.

Western Blotting Analysis: 4.0 µg/mL of this antibody detected DC-STAMP in 10 µg of mouse thymus lysate.

Target description

~58 kDa observed. Target band size appears larger than the calculated molecular weights of 53.39 kDa (human) and 53.88 kDa (mouse) due to glycosylation. Uncharacterized band(s) may appear in some lysates.

Physical form

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG2aκ antibody in PBS without preservatives.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.