Beta Hexosaminidase Activity Assay Kit

Produktnummer ABIN6253454

Produktdetails

Applikation: Activity Assay (AcA), Immunoassay (IA)

Proben: Plasma, Serum

Produktmerkmale: Beta Hexosamindase Activity Assay Kit is a simple fluorometric assay that measures beta hexosaminidase activity in a 96-well microtiter plate format. Each kit provides sufficient reagents to perform up to 100 assays, including blanks, beta hexosaminidase positive control and unknown samples.

Bestandteile:

1. Recombinant Beta Hexosaminidase : One 25 μL vial of a 50 μg/mL Recombinant Human Beta Hexosaminidase.
2. 10X Substrate : One 500 μL vial.
3. 5X Assay Buffer : One 30 mL bottle.
4. 10X Neutralization Buffer : One 30 mL bottle. 2

Anwendungsinformationen

Aufbereitung der Reagenzien:

• 1X Assay Buffer: Dilute the stock 5X Assay Buffer 1:5 with deionized water for a 1X solution. Stir or vortex to homogeneity. Store unused 1X Assay Buffer at room temperature.
• 1X Substrate: Dilute the 10X Substrate 1:10 with 1X Assay Buffer. For example, add 5 μL of 10X Substrate to 45 μL of 1X Assay Buffer for each well used. Note: Prepare only enough 1X Substrate for immediate use.
• 1X Neutralization Buffer: Dilute the stock 10X Neutralization Buffer 1:10 with deionized water for a 1X solution. Stir or vortex to homogeneity. Store unused 1X Neutralization Buffer at room temperature.

Testdurchführung:

1. Add 50 μL of Beta Hexosaminidase samples to the 96-well microtiter black plate.
2. Add 50 μL of the 1X Substrate to each well.
3. Incubate at 37 °C for 15 minutes protected from light.
4. Add 100 μL of the 1X Neutralization Buffer to each well.
5. Read the plate at an excitation wavelength of 365 nm and an emission wavelength 450 nm using a microplate fluorometer.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: -80 °C

Informationen zur Lagerung: Upon receipt, store the Recombinant Beta Hexosaminidase and 10X Substrate at -80°C. Store the remaining components at room temperature.

Antigendetails

Hintergrund: Hexosaminidase is an enzyme that hydrolyzes terminal N-acetyl-D-hexosamine residues from N-acetyl- β-D-hexosaminides. The enzyme, localized to cellular lysosomes, is a dimer formed from either an α or ß subunit which produces three possible dimer combinations. The αß combination (isozyme A) is the only combination with a known function: hydrolyzing GM2 ganglioside in vivo. The other two combinations of αα (isozyme B) and ßß (isozyme S) have been observed in cellular tissues but their function is unknown. Isozyme A works in combination with cofactor GM2 activator protein to degrade GM2 gangliosides and other molecules that have terminal N-acetyl hexosamines. Gene mutations that code for the ß subunit can result in Sandhoff disease. Mutations in the locus for the α subunit decrease the hydrolysis of GM2 gangliosides, which is the primary cause of Tay-Sachs disease. While both the α and ß subunits of lysosomal hexosaminidase can cleave GalNAc residues, only the α subunit is able to hydrolyze GM2 gangliosides because of a key residue, Arg-424, and a loop structure that forms from the amino acid sequence in the alpha subunit. The GM2 activator protein acts as a transporter to localize GM2 gangliosides to hexosaminidase, so a functional hexosaminidase enzyme is able to hydrolyze GM2 gangliosides into GM3 gangliosides.