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SMPD3 antibody (A04641) performs well in ELISA, WB and shows reactivity with Human.
- Validated for inter-lot consistency, specificity, and reproducibility
- Committed to quality since 1993
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Western blot analysis of SMPD3 using anti-SMPD3 antibody (A04641).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SMPD3 antigen affinity purified polyclonal antibody (Catalog # A04641) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SMPD3 at approximately 75 kDa. The expected band size for SMPD3 is at 71 kDa.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SMPD3 antigen affinity purified polyclonal antibody (Catalog # A04641) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SMPD3 at approximately 75 kDa. The expected band size for SMPD3 is at 71 kDa.
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Anti-SMPD3 Antibody Picoband®
SMPD3 antibody
| Reactive Species | Host | Application |
|---|---|---|
| Human | Rabbit | ELISA, WB |
Tested Applications
SMPD3 antibody was positively detected by WB in:
human K562 whole cell, human Caco-2 whole cell
Suggested blocking solution with 5% non-fat milk or BSA; (*)Recommended protein loading: 20-40 ug per lane
Recommend Dilution
Application
Dilution
Species
Western blot
0.25-0.5 μg/ml
Human
ELISA
0.1-0.5 μg/ml
-
* It is recommended to optimize the amount of this reagent in each experimental system to achieve the best results.
Reactive Data
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WB
ELISA
Human
Tested
Recommend
Monkey
Predicted
Predicted
Product Information
Product Name
Anti-SMPD3 Antibody Picoband®
View all SMPD3 Antibodies
View all SMPD3 Antibodies
Form
Lyophilized
Uniprot
Gene ID
Storage & Handling
At -20°C for one year from date of receipt.
After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months.
Avoid repeated freezing and thawing.
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
IgG
Immunogen
E.coli-derived human SMPD3 recombinant protein (Position: F184-D553). Human SMPD3 shares 88.9% amino acid (aa) sequence identity with both mouse and rat SMPD3.
Cross-Reactivity
No cross reactivity with other proteins.
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Observed Molecular Weight
75 kDa
Calculated Molecular Weight
71.1 kDa
Background of SMPD3
Sphingomyelin phosphodiesterase 3 is an enzyme that in humans is encoded by the SMPD3 gene. SMPD3 (Sphingomyelin phosphodiesterase 3; also Neutral sphingomyelinase 2/nSMase2) is a 69-74 kDa member of the neutral sphingomyelinase family of enzymes. It is a monomeric Golgi/plasma membrane enzyme that converts sphingomyelin (a plasma membrane lipid) into ceramide and phosphorylcholine. This generates second messenger components that participate in signal transduction. Human SMPD3 is a two transmembrane, 655 amino acids molecule. It contains an N-terminal luminal segment (aa 1-10), a cytoplasmic region (aa 32-64), and one catalytic domain (aa 340-646). Phosphorylation increases its MW to 78 kDa in SDS-PAGE. There is one potential isoform that possesses an Asn substitution for aa 569-587. Over aa 2-655, human SMPD3 shares 91% aa sequence identity with mouse SMPD3.
Cite This Product
Anti-SMPD3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A04641)
Antibody Homology Across Species
The results generated by this homology analysis tool for commonly used animal species are shown below. For unlisted species, please refer to the homology analysis between the immunogen sequence and the corresponding species.
A homology of ≥90% is recommended. Generally, higher homology indicates a greater potential for cross-species reactivity. However, protein expression levels may vary among different species, tissues, and cell types. Therefore, the homology analysis results are for reference only and do not guarantee antibody cross-reactivity or experimental performance.
Human
100%
High Homology
Monkey
99%
High Homology
Mouse
89%
Moderate Homology
Rat
89%
Moderate Homology
Dog
88%
Moderate Homology
Rabbit
88%
Moderate Homology
Cow
88%
Moderate Homology
Sheep
88%
Moderate Homology
Zebrafish
54%
Low homology
Homology ≥ 90%High Homology
70% ≤ Homology < 90%Moderate Homology
Homology < 70%Low homology
Protocols
Western Blot protocol for SMPD3 antibody
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Product Promise
We promise Anti-SMPD3 Antibody Picoband® performs excellently in relevant applications for ELISA, WB. If it doesn't perform as promised, we will refund the product. Learn more
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A04641
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$370.00
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This product is for Research Use Only. Not for use in diagnostic procedures.
Update date:
Oct 12, 2026