ASAH2 ELISA Kit

Catalog No. ABIN8055738

This Colorimetric ELISA kit is designed for the quantitative measurement of Human ASAH2.

Quick Overview for ASAH2 ELISA Kit (ABIN8055738)

Target: ASAH2 (N-Acylsphingosine Amidohydrolase (Non-Lysosomal Ceramidase) 2 (ASAH2))

Binding Specificity: AA 99-780

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5 pg/mL - 4000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma (EDTA), Plasma (heparin), Serum

Product Details ASAH2 ELISA Kit

Minimum Detection Limit: 62.5 pg/mL

Purpose: Sandwich Human ASAH2 ELISA Kit to quantitate Human ASAH2 in cell culture supernatants, serum and plasma (heparin, EDTA).

Analytical Method: Quantitative

Specificity: Expression system for standard: CHO
Immunogen sequence: F99-I780

Capture antibody and Detection antibody: a monoclonal antibody from mouse|a biotinylated polyclonal antibody from goat

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: < 10 pg/mL

Components:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Material not included: Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional). Pipettes and pipette tips capable of precisely dispensing 0.5 μL through 1 mL volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500 mL graduated cylinders. Test tubes for dilution.

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Sample Volume: 100 μL

Plate: Pre-coated

Protocol: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the target amount in sample captured in plate.

Assay Precision: Intra-Assay Precision (CV%): 7.1%, 6.9%, 6.5%
Inter-Assay Precision (CV%): 5.5%, 5.9%, 6.3%

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: 4 °C,-20 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Expiry Date: 12 months

Target Details for ASAH2

Target: ASAH2 (N-Acylsphingosine Amidohydrolase (Non-Lysosomal Ceramidase) 2 (ASAH2))

Alternative Name: ASAH2

Background:

Background: Neutral ceramidase is an enzyme that in humans is encoded by the ASAH2 gene. Ceramidases (EC 3.5.1.23), such as ASAH2, catalyze hydrolysis of the N-acyl linkage of ceramide, a second messenger in a variety of cellular events, to produce sphingosine. Sphingosine exerts both mitogenic and apoptosis-inducing activities, and its phosphorylated form functions as an intra- and intercellular second messenger.

Gene Full Name: N-acylsphingosine amidohydrolase 2

Gene ID: 56624

UniProt: Q9NR71