ASAH1 ELISA kit (Human)

SKU: SEC302Hu

Size: 96Tests

# of Times Cited in literature: 2

Prepare Time: 1-3 days(please inquire for mutiple units)

Target Name: ASAH1

Target Full Name: N-Acylsphingosine Amidohydrolase 1

Alternative Names: AC; ASAH; PHP; PHP32; Acid Ceramidase; Acylsphingosine deacylase; Putative 32 kDa heart protein

Target Species: Human

Uniprot: Q13510

Gene ID: 427

Featured Series: SE kit

Featured Series Function: Detects protein (regular version)

Specificity: Reactive with Human ASAH1 / N-Acylsphingosine Amidohydrolase 1

Method: Colormetric

Detection principle: Double-antibody Sandwich

Detection range: 1.56-100ng/mL

Sensitivity: 0.57ng/mL

Assay Time: 3h

Sample Size: 100uL

Recommended/Predicted Sample Types: Serum, Plasma, Tissue Homogenates, Cell Lysates, Cell Culture Supernates and other Biological Fluids

Assay Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%

Reproducibility test menthod: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level N-Acylsphingosine Amidohydrolase 1 (ASAH1) 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 1 (ASAH1) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100

Storage: 4°C for 1 month/ -20°C for long-term(One year within shelf life)

Shelf-life: 12 months

Specificity: This assay has high sensitivity and excellent specificity for detection of N-Acylsphingosine Amidohydrolase 1 (ASAH1). No significant cross-reactivity or interference between N-Acylsphingosine Amidohydrolase 1 (ASAH1) and analogues was observed.

Stability: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Assay procedure summary: 1. Prepare all reagents, samples and standards; 2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C; 3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C; 4. Aspirate and wash 3 times; 5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C; 6. Aspirate and wash 5 times; 7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C; 8. Add 50µL Stop Solution. Read at 450nm immediately.

Test 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 1 (ASAH1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to N-Acylsphingosine Amidohydrolase 1 (ASAH1). 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 1 (ASAH1), 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 1 (ASAH1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Research Area: Enzyme & Kinase;

References Citing This Product: The investigation of ceranib-2 on apoptosis and drug interaction with carboplatin in human non small cell lung cancer cells in vitro

Targeting Acid Ceramidase to Improve the Radiosensitivity of Rectal Cancer