SMPD1 Antibody (563418) [Unconjugated]

Western blot shows culture media of U‑87 MG human glioblastoma/astrocytoma parental cell line and SMPD1 knockdown U-87 MG cell line (KD). Nitrocellulose membrane was probed with Mouse Anti-Human SMPD1 Monoclonal Antibody (Catalog # MAB5348) followed by HRP-conjugated secondary antibody. A specific band was detected for SMPD1 at approximately 69.9 kDa (as indicated) in the parental U-87 MG cell line, but is significantly reduced in the knockdown U-87 MG cell line. Primary antibody concentration used: 2.5 µg/mL. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

U‑87 MG human glioblastoma/astrocytoma cell line culture medium were prepared and immunoprecipitation was performed using 2.0 μg of Mouse Anti-Human SMPD1 Monoclonal Antibody (Catalog # MAB5348) pre-coupled to Dynabeads Protein G. Immunoprecipitated SMPD1 was detected in Western Blot with MAB5348 used at 1/500. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

• Reactivity: Hu
• Applications: IP, B/N
• Clone: 563418
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

SMPD1 Antibody (563418) [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human SMPD1 isoform 1
  His62-Pro628
  Accession # NP_000534
• Specificity: Detects human SMPD1 in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human SMPD3 and recombinant mouse SMPD1 is observed.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: SMPD1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunoprecipitation 25 ug/mL
  • Neutralization 0.384 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for SMPD1 Antibody (563418) [Unconjugated]

• Acid sphingomyelinase
• aSMase
• ASMsphingomyelin phosphodiesterase
• EC 3.1.4.12
• NPD
• SMPD1
• sphingomyelin phosphodiesterase 1, acid lysosomal

Background

Sphingomyelin phosphodiesterase, also known as acid sphingomyelinase and encoded by the SMPD1 gene, is a lysosomal phosphodiesterase which belongs to the acid sphingomyelinase family (1). SMPD1 catalyzes the hydrolysis of sphingomyelin to ceramide and phosphorylcholine. Ceramide, a bioactive lipid, has emerged as an important signaling molecule involved in a variety of cellular processes such as cell differentiation, apoptosis, and proliferation (2). Activation of SMPD1 occurs by the removal, chemical modification or dimerization of its C-terminal cysteine residue (3). Deficiencies of SMPD1 result in a lysosomal storage disorder referred to as Niemann-Pick disease (4). rhSMPD1 was expressed without the last three C-terminal residues, and is therefore constitutively active.

1. Schuchman, E.H. et al. (1991) J. Biol. Chem. 266:8531.
2. Melendez, A.J. et al. (2008) Biochim. Biophys. Acta 1784:66.
3. Qiu, H. et al. (2003) J. Biol. Chem. 278:32744.
4. Smith, E.L. and Schuchman, E.H. (2008) FASEB J. 22:3419.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SMPD1 Antibody (MAB5348)(2)

Publications using MAB5348

• S Rana, C Espinosa-D, R Ruhl, N Chatterjee, C Hudson, E Fraile-Bet, A Agarwal, S Khou, CR Thomas, S Anand Differential regulation of microRNA-15a by radiation affects angiogenesis and tumor growth via modulation of acid sphingomyelinase Sci Rep, 2020-03-27;10(1):5581. 2020-03-27 [PMID: 32221387] (Western Blot, Mouse)
• Xiahui Xiong, Chia-Fang Lee, Wenjing Li, Jiekai Yu, Linyu Zhu, Yongsoon Kim et al. Acid Sphingomyelinase regulates the localization and trafficking of palmitoylated proteins Biology Open 2019-05-29 [PMID: 31142470]

Bioinformatics

• Gene Symbol: SMPD1
• Uniprot:
  • NP_000534()