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

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• ASAH2/N-acylsphingosine Amidohydrolase-2 Antibody

  Name: Anonymous

  Application: Western Blot

  Sample Tested: human primary hepatocytes

  Species: Human

  Verified Customer | Posted 12/18/2017

  Review for anti-ASAH2 antibody (NBP1-76360)

  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

  

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