nSMase Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Sphingomyelin is also present in all eukaryotic cell membranes, especially the plasma membrane, and is particularly concentrated in the nervous system because sphingomyelin is a major component of myelin, the fatty insulation wrapped around nerve cells by Schwann cells or oligodendrocytes. Multiple Sclerosis is a disease characterised by deterioration of the myelin sheath, leading to impairment of nervous conduction. nSMase (Neutral sphingomyelinase; Sphingomyelin phosphodiesterase 2) converts sphingomyelin to ceramide. Although widely expressed in all tissues examined, except the spleen, high enzymatic activity occurs only in the brain. Mice lacking Smpd2 and Smpd3 are completely devoid of neutral SMase activity.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-10865

• nSMase Antibodies
• nSMase ELISA Kits
• nSMase Lysate
• nSMase Proteins