ASAH2 ELISA Kit (Human) (OKCD01831)

Catalog No: OKCD01831 (Formerly GWB-KBBCU4)

Size:96WELLS

Human ASAH2 ELISA Kit Standard Curve

Product Info

• Predicted Species Reactivity: Homo sapiens|Human
• Application: Enzyme-linked Immunosorbent assay-Sandwich
• ELISA Kit Detection Method: Colorimetric
• ELISA Kit Duration: 3h
• ELISA Kit Linearity:

  Sample	1:2 Dilution	1:4 Dilution	1:8 Dilution	1:16 Dilution
  Serum (n=5)	90-102%	96-105%	79-89%	81-94%
  EDTA Plasma (n=5)	90-98%	81-95%	92-99%	78-93%
  Heparin Plasma (n=5)	94-105%	80-88%	98-105%	81-104%

• ELISA Kit Principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to N-Acylsphingosine Amidohydrolase 2 (ASAH2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to N-Acylsphingosine Amidohydrolase 2 (ASAH2). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain N-Acylsphingosine Amidohydrolase 2 (ASAH2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of N-Acylsphingosine Amidohydrolase 2 (ASAH2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
• ELISA Kit Range: 0.78-50ng/mL
• ELISA Kit Recovery:

  Matrix	Recovery Range (%)	Average (%)
  Serum (n=5)	97-105	101
  EDTA Plasma (n=5)	82-89	85
  Heparin Plasma (n=5)	80-101	85

• ELISA Kit Reproducibility: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level N-Acylsphingosine Amidohydrolase 2 (ASAH2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level N-Acylsphingosine Amidohydrolase 2 (ASAH2) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
• ELISA Kit Component:
  • Anti-ASAH2 Microplate: 96 Wells (12 x 8 Well Strips)
  • ASAH2 Lyophilized Standard: 2 x 50 ng
  • 100X Biotinylated ASAH2 Detector Antibody: 120 uL
  • Avidin/HRP Conjugate: 120 uL
  • Standard Diluent: 1 x 20 mL
  • Detector Antibody Diluent: 1 x 12 mL
  • Conjugate Diluent: 1 x 12 mL
  • 30X Wash Buffer: 1 x 20 mL
  • TMB Substrate: 1 x 9 mL
  • Stop Solution: 1 x 6 mL
• Reconstitution and Storage: 2°C to 8°C|-20°C
• Sample Type: Serum, plasma, tissue homogenates and other biological fluids
• Sensitivity: < 0.31 ng/mL
• Specificity: This assay has high sensitivity and excellent specificity for detection of N-Acylsphingosine Amidohydrolase 2 (ASAH2). No significant cross-reactivity or interference between N-Acylsphingosine Amidohydrolase 2 (ASAH2) and analogues was observed.
• Assay Info: Assay Methodology: Quantitative Sandwich Immunoassay

Target Info

• Gene Symbol: ASAH2
• Gene Full Name: N-acylsphingosine amidohydrolase 2
• Alias Symbols: acylsphingosine deacylase 2;BCDase;HNAC1;LCDase;mitochondrial ceramidase;N-acylsphingosine amidohydrolase (non-lysosomal ceramidase) 2;N-acylsphingosine amidohydrolase 2;NCDase;N-CDase;neutral ceramidase;neutral/alkaline ceramidase;non-lysosomal ceramidase.
• NCBI Gene Id: 56624
• Protein Name: Neutral ceramidase
• Description of Target: Hydrolyzes the sphingolipid ceramide into sphingosine and free fatty acid at an optimal pH of 6.5-8.5 (PubMed:10781606, PubMed:26190575). Acts as a key regulator of sphingolipid signaling metabolites by generating sphingosine at the cell surface. Acts as a repressor of apoptosis both by reducing C16-ceramide, thereby preventing ceramide-induced apoptosis, and generating sphingosine, a precursor of the antiapoptotic factor sphingosine 1-phosphate. Probably involved in the digestion of dietary sphingolipids in intestine by acting as a key enzyme for the catabolism of dietary sphingolipids and regulating the levels of bioactive sphingolipid metabolites in the intestinal tract.
• Uniprot ID: Q9NR71
• Protein Accession #: NP_001137446.1
• Nucleotide Accession #: NM_001143974.1
• Protein Size (# AA): 780