Preparing Amyloid beta oligomer - vehicle toxicity...... (Mar/12/2011 )
greeting of the day,
I am stuck with one experment. My drug vehicle is showing toxicity (About 50 %)and i am not able figure out the reason behind this.
Acc. to protocol i prepare the drug in this way,(vehicle and drug prepared in the same way)
Drug----->dissolved in HFIP(Solvent)-----> HFIP is evaporated in Fume hood for 24 h-----
>then stock (5mM) is prepared by DMSO------> futher 100 uM is prepared by PBS.
Now this is used for MTT done on HT-22 cells (Hippocampal neuron).
I used 3 conc-->2uMm, 5uM and 10uM. There is increase in toxicity in 2uM and 5uM. And to my surprize, in 10uM vehicle the viability has incresed in comparison to 2uM which seems to be quite impossible. i am worried and tense and not able to figure out the reason behind.
I just wanted to know from the people here that has similar happened withanyone.
Any kind of advice or views will be highly appreciated.
Thanx in advance
I would be glad to know about the people who are working with amyloid Beta oligomer. And basically i am interseted in knowing their method of preparation of oligomers.