Acid sphingomyelinase Recombinant Rabbit Monoclonal Antibody [PSH15-61]
Catalog# HA723768
Acid sphingomyelinase Recombinant Rabbit Monoclonal Antibody [PSH15-61]
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WB
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IHC-P
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IP
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Human
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Mouse
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Rat
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Green monkey
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HA751572
不含抗保成分
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unconjugated
Overview
Product Name
Acid sphingomyelinase Recombinant Rabbit Monoclonal Antibody [PSH15-61]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human Acid sphingomyelinase aa 47-631.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IHC-P, IP
Molecular Weight
Predicted band size: 70 kDa
Positive Control
THP-1 treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300n/mL BFA for 3 hours cell lysate, HepG2 cell lysate, HepG2 treated with 10ng/mL BFA for 1.5 hours cell lysate, COS-1 cell lysate, PC-12 cell lysate, PC-12 starved for 16 hours then treated with 200nM TPA for 4 hours cell lysate, RAW264.7 cell lysates, human kidney tissue, human liver tissue, mouse kidney tissue, mouse liver tissue, rat kidney tissue, rat liver tissue.
Conjugation
unconjugated
Clone Number
PSH15-61
Reactivity Data
| WB | IHC-P | IP | |
|---|---|---|---|
| Human |
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|
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| Mouse |
|
|
|
| Rat |
|
|
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| Green Monkey |
|
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000-1:5,000
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IHC-P
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1:100-1:200
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IP
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1-2μg/sample
Target
Function
Acid sphingomyelinase is one of the enzymes that make up the sphingomyelinase (SMase) family, responsible for catalyzing the breakdown of sphingomyelin to ceramide and phosphorylcholine. They are organized into alkaline, neutral, and acidic SMase depending on the pH in which their enzymatic activity is optimal. Acid sphingomyelinases' (aSMases) enzymatic activity can be influenced by drugs, lipids, cations, pH, redox and other proteins in the environment. Specifically aSMases have been shown to have increased enzymatic activity in lysobisphosphatidic acid (LBPA) or phosphatidylinositol (PI) enriched environments, and inhibited activity when phosphorylated derivatives of PI are present. Sphingomyelin phosphodiesterase 1 (SMPD1) is the gene that codes for two aSMase enzymes distinct in the pools of sphingomyelin they hydrolyse. Lysosomal sphingomyelinase (L-SMase) is found in the lysosomal compartment, and the secretory sphingomyelinase (S-SMase) is found extracellularly.
Background References
1. Pinto C et al. Acid Sphingomyelinase Deficiency: A Clinical and Immunological Perspective. Int J Mol Sci. 2021 Nov
2. Mauhin W et al. Acid Sphingomyelinase Deficiency: Sharing Experience of Disease Monitoring and Severity in France. J Clin Med. 2022 Feb
Subcellular Location
Lysosome, Lipid droplet, Secreted.
Synonyms
Acid sphingomyelinase antibody
ASM antibody
ASM_HUMAN antibody
aSMase antibody
NPD antibody
Smpd1 antibody
Sphingomyelin phosphodiesterase 1 acid lysosomal antibody
Sphingomyelin phosphodiesterase antibody
Images
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☑ Cell treatment (CT)
Western blot analysis of Acid sphingomyelinase on different lysates with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/5,000 dilution.
Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: THP-1 treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300n/mL BFA for 3 hours cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: HepG2 treated with 10ng/mL BFA for 1.5 hours cell lysate (20 µg/Lane)
Lane 5: MOLT-4 cell lysate (low expression) (20 µg/Lane)
Lane 6: COS-1 cell lysate (20 µg/Lane)
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 25 seconds; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723768) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Western blot analysis of Acid sphingomyelinase on different lysates with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/5,000 dilution.
Lane 1: PC-12 cell lysate (40 µg/Lane)
Lane 2: PC-12 starved for 16 hours then treated with 200nM TPA for 4 hours cell lysate (40 µg/Lane)
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 1 minute 18 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723768) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Acid sphingomyelinase on RAW264.7 cell lysates with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/2,000 dilution.
Lysates/proteins at 30 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723768) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723768) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723768) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723768) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723768) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723768) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Acid sphingomyelinase antibody (HA723768) at 1/100 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723768) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Acid sphingomyelinase was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA723768 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723768 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HepG2 cell lysate (input)
Lane 2: HA723768 IP in HepG2 cell lysate
Lane 3: Rabbit IgG instead of HA723768 in HepG2 cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 46 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"