Hi there,

There are some commercial kits based on the coupled fluorometric detection of aspartate:

https://www.abcam.com/asparaginase-activity-assay-kit-ab107922.html

https://www.sigmaaldrich.com/catalog/product/SIGMA/MAK007?lang=fr®ion=FR

Hello Basem, I adopt the Sigma-Aldrich protocol for L-asparaginase activity assay Kit

Aspartate Standards for Colorimetric Detection Dilute 10 μL of the 100 mM aspartate standard solution with 990 μL of Asparaginase Assay Buffer to prepare a 1 mM standard solution. Add 0, 2, 4, 6, 8, and 10 μL of the 1 mM Aspartate standard solution into a 96 well plate, generating 0 (blank), 2, 4, 6, 8, and 10 nmole/well standards. Add Asparaginase Assay Buffer to each well to bring the volume to 50 μL.

Aspartate Standards for Fluorometric Detection Dilute 10 μL of the 100 mM (100 nmole/μL) Aspartate Standard Solution with 990 μL of Asparaginase Assay Buffer to prepare a 1 mM (1 nmole/μL) standard solution. Dilute 10 μL of the 1 mM standard solution with 90 μL of Asparaginase Assay Buffer to generate a 0.1 mM (0.1 nmole/μL) standard solution. Add 0, 2, 4, 6, 8, and 10 μL of the 0.1 mM Asparate standard solution into a 96 well plate, generating 0 (blank), 0.2, 0.4, 0.6, 0.8, and 1.0 nmole/well standards. Add Asparaginase Assay Buffer to each well to bring the volume to 50 μL.

Details especially calculations and expressions and more could be found in the attached documents.

Thank you for the answers but actually i have been looking for a manual techniques actually that would be more cheaper than the kit.. Although thank you very much for the help

This assay for asparaginase relies on the conversion of NADP to NAD+ as measured spectrophotometrically. It is designed for use in 96-well plates and for measuring asparaginase in serum but having a look at the paper I see absolutely no reason it wouldn't work in cuvettes with larger volumes and with a variety of sample types. Materials required are inexpensive.

Best of luck! Let me know if it works out!

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