Active Recombinant Human SMPD1 protein, His-tagged

• Cat.No.: SMPD1-656H
• Product Overview: Active Recombinant Human SMPD1 protein(NP_000534.3) (Met 1-Cys 631) was expressed in Insect Cells, fused with a polyhistidine tag at the C-terminus.

Specification

• Species: Human
• Source: Insect Cells
• Tag: His
• Protein Length: 1-631 aa
• Form: Lyophilized from sterile 50mM Tris, 100mM NaCl, pH 8.0, 0.1% OGP, 10% glycerol.
• Bio-activity: Measured by its ability to cleave. 2-N-Hexadecanoylamino-4-nitrophenylphosphorylcholine (HNPPC). The specific activity is >1,000 pmol/min/μg.
• Molecular Mass: The secreted recombinant human SMPD1 consists of 518 amino acids and predicts a molecular mass of 65 kDa as estimated by SDS-PAGE under reducing conditions.
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: > 94 % as determined by SDS-PAGE
• Storage: Store it under sterile conditions at -20°C to -80°C upon receiving. Recommend to aliquot the protein into smaller quantities for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.
• protein Refseq-Weblink: http://www.ncbi.nlm.nih.gov/protein/NP_000534.3
• Unit ID: P17405
• Unit ID-Weblink: https://www.uniprot.org/uniprotkb/P17405
• Publications:
  • Essential role for acid sphingomyelinase-inhibited autophagy in melanoma response to cisplatin (2016)

Gene Information

• Gene Name: SMPD1 sphingomyelin phosphodiesterase 1, acid lysosomal [ Homo sapiens ]
• Official Symbol: SMPD1
• Synonyms: SMPD1; sphingomyelin phosphodiesterase 1, acid lysosomal; sphingomyelin phosphodiesterase; acid sphingomyelinase; ASM; NPD; ASMASE
• Gene ID: 6609
• mRNA Refseq: NM_000543
• protein Refseq: NP_000534
• MIM: 607608

Citation

Essential role for acid sphingomyelinase-inhibited autophagy in melanoma response to cisplatin

Journal: Oncotarget    PubMed ID: 27107419    Data: 2016/5/3

Authors: Davide Cervia, Emma Assi, Cristiana Perrotta

Article Snippet:Cisplatin (Cisplatino Teva) was from Teva Pharma Italia (Milano, Italy).Cisplatin (Cisplatino Teva) was from Teva Pharma Italia (Milano, Italy).. Human A-SMase (Recombinant Human SMPD1, His-tagged) was purchased from Creative BioMart (Shirley, NY, USA).. Annexin V-Fluorescein Isothiocianate (FITC) and PI were obtained from Life Technologies (Monza, Italy) and eBioscience (San Diego, CA, USA), respectively.Annexin V-Fluorescein Isothiocianate (FITC) and PI were obtained from Life Technologies (Monza, Italy) and eBioscience (San Diego, CA, USA), respectively.

A. Dose-response curves of the effects of cisplatin on the viability of B16-W6_scr and B1-W6_pSIL10, as measured by the MTT assay ( n = 10). B. Measurement of A-SMase activity in B16-W6_scr and B1-W6_pSIL10 cells treated with cisplatin (10 μg/ml, 16 h). Activity was expressed as fold increase of untreated cells ( n = 5). Statistical significance * p < 0.05 vs untreated B16-W6_scr. C. B1-W6_pSIL10 and B16-W6_scr were cultured for 16 h in the absence (UT: untreated) or in the presence of cisplatin (10 μg/ml). B1-W6_pSIL10 and B16-W6_scr were also treated with human A-SMase (2.0 units/ml) or amitriptyline (5 μM, 1 h before cisplatin treatment), respectively. Left panels: representative dot plots of Annexin V-FITC/PI staining; Right panel: apoptosis quantification expressed as fold increase of total apoptotic cells (Annexin V + /PI ? and Annexin V + /PI + cells) compared to their respective UT controls ( n = 8). Statistical significance *and § p < 0.05 vs B1-W6_pSIL10 and B16-W6_scr, respectively. D. Western blot analysis of cleaved Caspase 3 expression in B16-W6_scr cells cultured for 16 h in the absence (UT) or in the presence of cisplatin (10 μg/ml) and cisplatin + amitriptyline (5 μM, 1 h before cisplatin treatment). GAPDH was used as the internal standard. The images are representative of results obtained from three experiments.