Anti-V-ATPase subunit d 2

V-type proton ATPase subunit d 2 (UniProt Q80SY3; also known as Osteoclast-specific vacuolar ATP synthase, V-ATPase subunit d 2, Vacuolar proton pump subunit d 2) is encoded by the Atp6v0d2 gene (Gene ID 242341) in murine species. V-ATPase is a proton pump consists of an 8-subunit (subunit A through H) peripheral catalytic V1 complex (V1) and a 6-subunit (a, c, c , d, e, and c′ in yeast or Ac45 in higher eukaryotes) catalytic integral membrane V0 proton pore complex. V0 subunit d is a 38 kDa protein located at the cytosolic face of the membrane. There exist two mammalian subunit d isoforms, d1 and d2, encoded by two separate genes, ATP6V0D1 and ATP6V0D1. While d1 is expressed ubiquitously, d2 was expressed in specific tissues, such as kidney, lung, and bone. In addtion, d2 expression level is about 5-fold higher than d1 in osteoclasts. Atp6v0d1-knockout is embryonic lethal in mice, while Atp6v0d2 deletion does not affect V-ATPase function, but decreases cell to cell fusion. V-ATPase subunit d 2 transcription is mediated by transcription factor nuclear factor of activated T cells cytoplasmic 1 (NFATc1) that is essential for osteoclastogenesis.

~38 kDa observed. 40.43/40.49/40.52 kDa (human/mouse/rat) calculated.

Epitope: C-terminus

Linear peptide corresponding to human/mouse/rat V-ATPase subunit d 2 C-terminal end sequence.

Detect using this rabbit polyclonal Anti-V-ATPase subunit d 2 Antibody, Cat. No. ABS1677, validated for use in Western Blotting.

Research Category
Signaling

Western Blotting Analysis: A representative lot detected higher V-ATPase subunit d 2 expression in osteoclasts differentiated in culture from murine bone marrow monocyte precursors (BMMs) infected by RACK1-expressing retrovirus due to an upregulated NFATc1 expression caused by RACK1-overexpression (Lin, J., et al. (2015). Sci. Signal. 8(379):ra54).

Western Blotting Analysis: A representative lot detected less V-ATPase subunit d 2 expression in osteoclasts differentiated in culture from GSK3 -S9A Tg mouse bone marrow monocyte precursors (BMMs) due to a downregulated NFATc1 expression caused by osteoclast-specific GSK3 -S9A mutant expression (Jang, H.D., et a1. (2011). J. Biol. Chem. 286(45):39043-39050).

Western Blotting Analysis: A representative lot detected V-ATPase subunit d 2 in osteoclasts, but not bone marrow monocyte precursors (BMMs), from wild-type mice, nor in osteoclasts from Atp6v0d2-knock mice (Lee, S.H., et al. (2006). Nat. Med. 12(12):1403-1409).

Broad species reactivity expected based on high sequence homology.

This polyclonal antiserum was raised against V-ATPase subunit d 2 C-terminal end sequence conserved among human, mouse, and rat species. Specificity was confirmed by the lack of cross-reactivity to mouse Atp6v0d1 overexpressed in 293 cells (Lee, S.H., et al. (2006). Nat. Med. 12(12):1403-1409).

Format: Unpurified

Rabbit polyclonal antibody serum with 0.05% sodium azide.

Unpurified.

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.

Evaluated by Western Blotting in mouse osteoclasts lysate.

Western Blotting Analysis: A 1:2,000 dilution of this antibody detected V-ATPase subunit d 2 in mouse osteoclast lysate.

Concentration: Please refer to lot specific datasheet.

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