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.

Detection of SMPD1 by Immunoprecipitation.

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).

Applications for Human SMPD1 Antibody

Application

Recommended Usage

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human SMPD1
(Catalog # 5348-PD), see our available Western blot detection antibodies.

Western Blot

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.

Neutralization

Measured by its ability to neutralize Recombinant Human SMPD1 (0.5 µg/mL, Catalog # 5348-PD) cleavage of the substrate 2-N-Hexadecanoylamino- 4-nitrophenylphosphorylcholine (HNPPC, 250 µM). The Neutralization Dose (ND₅₀) is typically 3.0 µg/mL

Reviewed Applications

Read 1 review rated 5 using MAB5348 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Background: SMPD1

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.

References

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

Long Name

Sphingomyelin Phosphodiesterase 1, Acid Lysosomal

Alternate Names

aSMase, NPD

Entrez Gene IDs

6609 (Human); 20597 (Mouse); 308909 (Rat)

Gene Symbol

SMPD1

UniProt

NP_000534

Additional SMPD1 Products

Product Documents for Human SMPD1 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human SMPD1 Antibody

For research use only

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