ASAH2/N-acylsphingosine Amidohydrolase-2 Antibody - BSA Free

Western Blot: ASAH2/N-acylsphingosine Amidohydrolase-2 Antibody [NBP1-76360] - ASAH2 Antibody 293 cell lysate with ASAHL antibody at (A) 1 and (B) 2 ug/ml.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ELISA
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

ASAH2/N-acylsphingosine Amidohydrolase-2 Antibody - BSA Free Summary

• Immunogen: Antibody was raised against a 17 amino acid synthetic peptide near the carboxy terminus of the human ASAH2. The immunogen is located within the last 50 amino acids of ASAH2. Amino Acid Squence: YRIRYFGHNRKQDILK
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ASAH2
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Western Blot 1-2 ug/ml
• Control Peptide:

  ASAH2/N-acylsphingosine Amidohydrolase-2 Peptide (NBP1-76360PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Peptide affinity purified

Alternate Names for ASAH2/N-acylsphingosine Amidohydrolase-2 Antibody - BSA Free

• ASAH2 N-acylsphingosine amidohydrolase (non-lysosomal ceramidase) 2
• ASAH2
• BCDase
• HNAC1
• LCDase
• Nacylsphingosine Amidohydrolase2
• NCDase
• N-CDase

Background

Sphingolipids are hydrolyzed by ceramidases to yield sphingosine and fatty acids. These ceramidases are classified according to the pH range that supports their optimal activity. ASAH2 is a neutral ceramidase and key regulator of sphingolipid signaling metabolites at the cell surface, catalyzing the hydrolysis of the N-acyl linkage of ceramide at an optimal pH of 6.5-8.5. ASAH2 is a type II integral membrane protein that can be cleaved to yield a soluble secreted protein and acts as a repressor of apoptosis both by reducing C16-ceramide, thereby preventing ceramide-induced apoptosis, and generating sphingosine. Sphingosine exerts both mitogenic and apoptosis-inducing activities, and its phosphorylated form functions as an intra- and intercellular second messenger. ASAH2 is ubiquitously expressed primarily expressed with higher levels in the intestine, kidney, skeletal muscle and heart. Recent studies indicate that ASAH2 encoded neutral ceramidase is a key enzyme for the catabolism of dietary sphingolipids and regulates the levels of bioactive sphingolipid metabolites in the intestinal tract.

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.

Review for ASAH2/N-acylsphingosine Amidohydrolase-2 Antibody (NBP1-76360) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 12/18/2017

Summary

• Application: Western Blot
• Sample Tested: human primary hepatocytes
• Species: Human
• Lot: 4743-0803

Comments

• Comments: Name: Anti-ASAH2 antibody (NBP1-76360)
  Catalog #: Anti-ASAH2 antibody (NBP1-76360)
  Lot Number: Anti-ASAH2 antibody (NBP1-76360, Lot # 4743-0803)
  PO/Order Number: Click here to enter text..
  WB Image Description (Please provide labels for all lanes): lane 1: human primary hepatocytes; lane 2: Hep G2
  Sample Information:
       Cell Line or Tissue: human primary hepatocytes; and Hep G2
       Species: human
       Treatment: No treatment
  Lysate Preparation:
       Date of lysate preparation: December 4, 2017
       Lysis buffer used: 1X lysis buffer from Cell Signaling by adding PMSF
  Reducing agent: beta-mercaptoethanol, DTT
  If boiled (temperature/time): Yes
  Controls:
       Positive Control: No
       Negative Control: No
       Loading Control (please attach additional images if applicable): No
  Protein Amount Loaded per lane: 20 ug
  Antibody Storage Conditions: -20℃
  Electrophoresis:
  Gel Percentage: 10%
  Electrophoresis Conditions: Tris-Glycine-SDS at room temperature
       Voltage: 120V
       Time: 2 hours
  Membrane Transfer:
       Method (Submersion/Semi-dry): wet transfer
       Membrane Type (PVDF/Nitrocellulose): Nitrocellulose
       Time: 2 hours
       Voltage: 100V
  Blocking:
       Blocking Solution: 5% milk in 1X TBST
       Time: 1 hour at room temperature
  Primary Antibody:
       Dilution: 1/1000
       Diluent Buffer: 2.5% BSA
       Incubation Time: overnight
       Incubation Temperature: 4℃
  Washing Conditions:
       Wash Solution: 1X TBST
       Time and Repetitions: 5 min each for 3 times
  Secondary Antibody
  Manufacturer and Catalog #: Promega, W401B, Lot # 0000187662
  Secondary description: goat anti-rabbit secondary antibody
       Dilution: 1/2000
       Diluent Buffer: 3% milk
       Incubation Time: 1 hour
       Incubation Temperature: room temperature
  
  Detection Method:
  Detection: ECL (GE, cat # RPN2209, lot # 9838243)
  Procedure: Add equal volume of A and B, mix and apply on the membranes for 3-5 min before exposure
  Development Time: 45 seconds
  Molecular weight of band(s): ~ 70 kDa
  Experimental Concerns and Observations: A specific band around 70 kDa was observed

Bioinformatics

• Gene Symbol: ASAH2
• Uniprot:
  • NP_063946(Human)