Recombinant Human Neutral ceramidase (ASAH2), partial

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Code CSB-EP865115HU
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Size $306
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Product Details

Purity
Greater than 85% as determined by SDS-PAGE.
Target Names
ASAH2
Uniprot No.
Research Area
Signal Transduction
Alternative Names
N-CDase;NCDase;Acylsphingosine deacylase 2;BCDase;LCDase;hCD;N-acylsphingosine amidohydrolase 2;Non-lysosomal ceramidase
Species
Homo sapiens (Human)
Source
E.coli
Expression Region
610-780aa
Target Protein Sequence
FRNLAKAIATDTVANLSRGPEPPFFKQLIVPLIPSIVDRAPKGRTFGDVLQPAKPEYRVGEVAEVIFVGANPKNSVQNQTHQTFLTVEKYEATSTSWQIVCNDASWETRFYWHKGLLGLSNATVEWHIPDTAQPGIYRIRYFGHNRKQDILKPAVILSFEGTSPAFEVVTI
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Mol. Weight
25.2 kDa
Protein Length
Partial
Tag Info
N-terminal 6xHis-tagged
Form
Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
3-7 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA
Please contact us to get it.
Description

The process for producing recombinant Human ASAH2 protein in e.coli cells follows a sequence of steps: creating the expression vector that codes for the Human ASAH2 protein (610-780aa), introducing it into e.coli cells, cultivating and inducing of positive cells for protein expression, lysing cells, and conducting expression analysis. The resultant recombinant Human ASAH2 protein is purified from the cell lysate using affinity purification, with its purity exceeding 85%, as determined by SDS-PAGE.

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Target Background

Function
Plasma membrane ceramidase that hydrolyzes sphingolipid ceramides into sphingosine and free fatty acids at neutral pH. Ceramides, sphingosine, and its phosphorylated form sphingosine-1-phosphate are bioactive lipids that mediate cellular signaling pathways regulating several biological processes including cell proliferation, apoptosis and differentiation. Also catalyzes the reverse reaction allowing the synthesis of ceramides from fatty acids and sphingosine. Together with sphingomyelinase, participates in the production of sphingosine and sphingosine-1-phosphate from the degradation of sphingomyelin, a sphingolipid enriched in the plasma membrane of cells. Also participates in the hydrolysis of ceramides from the extracellular milieu allowing the production of sphingosine-1-phosphate inside and outside cells. This is the case for instance with the digestion of dietary sphingolipids in the intestinal tract.
Gene References into Functions
  1. The structural basis for ceramide recognition and hydrolysis by human neutral ceramidase has been uncovered. PMID: 26190575
  2. These results demonstrate for the first time that adiponectin inhibits TNFalpha-induced inflammatory response via Cav1-mediated ceramidase recruitment and activation in an AdipoR1-dependent fashion. PMID: 24397980
  3. cloned a 3000 bp region upstream of the translational start site of the nCDase gene PMID: 21531200
  4. c-Jun/AP-1 signaling may, in part, regulate serum-induced nCDase gene transcription PMID: 21530485
  5. Both acidic ceramidase (aCDase) and neutral ceramidase (nCDase) activities declined after low- and high-UVB, but returned to normal only in low-UVB cells PMID: 20520628
  6. Blocking acid ceramidase but not sphingosine kinase activity in alveolar macrophages led to decreased ERK and Akt activity and induction of cell death. PMID: 15210766
  7. Neutral ceramidase is localized mainly in plasma membranes. PMID: 15845354
  8. the present study identified a novel amidase sequence containing a critical serine residue that may function as a nucleophile in the hydrolytic attack on the amide bond present in ceramide PMID: 16229686
  9. Important for the generation of sphgosine 1 phosphqte (S1P) and S1P-mediated cell proliferation and survival. PMID: 16940153
  10. the purification and characterization of neutral ceramidase from human ileostomy content, using octanoyl-[(14)C]sphingosine as substrate PMID: 17475390
  11. serine palmitoyltransferase and ceramidase are two major ceramide metabolizing enzymes that may have roles in psoriatic epidermis PMID: 17982236

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Subcellular Location
[Neutral ceramidase]: Cell membrane; Single-pass type II membrane protein. Membrane raft; Single-pass type II membrane protein. Membrane, caveola; Single-pass type II membrane protein. Golgi apparatus membrane; Single-pass type II membrane protein. Mitochondrion. Secreted, extracellular exosome.; [Neutral ceramidase soluble form]: Secreted.
Protein Families
Neutral ceramidase family
Tissue Specificity
Primarily expressed in intestine. Ubiquitously expressed with higher levels in kidney, skeletal muscle and heart. The ubiquitous expression observed for ASAH2 might be an experimental artifact due to the paralog ASAH2B.
Database Links

HGNC: 18860

OMIM: 611202

KEGG: hsa:56624

STRING: 9606.ENSP00000378897

UniGene: Hs.512645

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