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Technical Data Sheet

SP4061

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nSMase2 (C-terminal region)
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
72 kDa

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Western blot of adult mouse brain. The blots were probed with anti-nSMase2 (C-terminal region) rabbit polyclonal antibody at 1:2000 (lane 1), 1:1000 (lane 2), or 1:500 in the absence (lane 3) or presence of nSMase2 blocking peptide (SX4065) (lane 4).

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Enlarge

Immunocytochemical labeling of nSMase2 in aldehyde-fixed and NP-40-permeabilized differentiated PC12 cells. The cells were labeled with rabbit polyclonal anti-nSMase2 (SP4061) antibody in the absence (Left) or presence (Right) of blocking peptide (SX4065). The antibody was detected using appropriate secondary antibody conjugated to DyLight® 594. The cells were counterstained with Sytox green to label nuclei.

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Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Moylan, J. et al. (2014) Redox biology (2)910: 920. WB: C2C12 myoblast
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Background
Cellular stress leads to hydrolysis of sphingomyelin to generate lipid second messenger molecules including ceramide, sphingosine, and sphingosine-1-phosphate. A variety of sphingomyelinase activities have been described that differ in tissue and subcellular distribution, as well as pH and cation dependance. These enzymes generate ceramide for specific signaling pathways that lead to a wide range of cellular responses, such as apoptosis, cell cycle arrest, cell survival, and cell proliferation. Neutral sphingomyelinases (nSMases) are Mg2+-dependent neutral pH SMases, and the family includes nSMase1, nSMase2, and nSMase3. These nSMases differ in their tissue distribution and subcellular localization. nSMase2 (also known as SMPD3) is expressed primarily in brain, and co-localizes with Golgi markers in several cell lines. The activity and phosphorylation of nSMase2 is upregulated in response to oxidative stress. In addition, nSMase2 binds calcineurin, and this interaction inhibits nSMase2 activity.


Background References
Hofmann, K. et al. (2000) Proc Natl Acad Sci U S A. 97(11):5895.
Stoffel, W. et al. (2005) Proc Natl Acad Sci U S A. 102(12):4554.
Filosto, S. et al. (2010) J Biol Chem. 285(14):10213.
Immunogen
nSMase2 synthetic peptide corresponds to amino acids in the C-terminal region of human nSMase2. This sequence is highly conserved in rat and mouse nSMase2, and has low homology to other nSMase family members.
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
The antibody detects a 78 kDa* band corresponding to nSMase2 on SDS-PAGE immunoblots of adult mouse brain and rat PC12 cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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