Has anyone attempted to follow the protocol by Yamamoto Transitional Oncology vol 1 number 2 July 2008 pp 65-72. The assay claims to measure Nagalase an enzyme secreted from cancer cells. We have followed the published protocol without success and would appreciate it if anyone who has had experience would contact us.
Did you try a positive control enzyme, alpha-galactosidase? It is available from Sigma. That should help you decide whether the problem is with the sample or the detection.
Sorry Onur. My interest at present is aimed at measuring tumor burden reduction, hence my query re the measurement of the Nagalase enzyme.
There are a few references to the preparation of GcMAF in the Yamamoto papers, however to date I have not attempted to examine the methodology given.
Ta, Bill McKay
Still no news? We are also having issues with nagalase determination.
I have tested nagalase concentration in few sample. It was not hopeful.
But I would like to try again!
Which method have you tried? What elements didn't work out in the test? We are trying a capture method on a plate. Afterwards we do an incubation with substrate and follow the reaction kinetics. We first tried with 4-Nitrophenyl N-acetyl-α-D-galactosaminide, but this gave just gave noise signal. No real influence. Then we tried 4MU-alpha-N-acetyl-D-galactosaminide. Normally the fluorescent signal should grow in function of time, but we see the opposite. But it seems to correlate a bit with nagalase concentration.
- Badges
- Science topic
- Similar topics
- Medicine
- Oncology
- Cancer Research