Hi. I'm working on BV2 cells. I'm having problem in NO production after LPS induction. I'm using 10μg/ml of LPS concentration and incubate it for 24 hours. . Sometimes the production of NO will be five fold when compared to control and sometimes no or low production. The same problem even when we use new batch of cells. Is the NO production got to do with usage of colorless media with L-glutamine? Please do help me. Thank you.
Hello Careena,
To me, 10 ug/ml is a quite high concentration of LPS which might cause this. You could use 1 ug/ml and see whether it is consistent or not. The number of cells also affect the level of NO. You also should be consistent with the same cell number. BV2 cells are sometimes self inflammatory when they become over confluent. So, keep your eyes that your cells are not over confluent at the end of 24 h incubation with LPS.
With best
Hello Mr. Moniruzzam,
Thank you for your opinion. The problem that we are facing is sometimes there is NO production and the next time there will have no NO production.
Please consider that iNOS might need extracellular L-arginine supply. YOu probably get a more stable effect when supplementing your buffer with L-arginine. Hope this helps. By the way, there is a nice fluorescence NO sensor offered by NGFI (www.ngfi.eu).
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