I would like to know how to further dilute it to apply on cancer cell line for cytotoxicity assay cck8
The previously mentioned solvent was recommended by the supplier sigmaaldrich
Dear Mme./Ms/Mrs. Hayat Youssef,
Honestly spoken:
think practically.... (unfortunately you have not informed about the amount (grams or ml) of 'monophosphoryl lipid' just to comment also on how much of the solvent "dilution" to get a certain percentage of the substrate IN the solvent for your cytotoxicity assay).
i) you have solved a /your substrate ['monophosphoryl lipid'] in a mixture of
- Chloroform: 74 (e. g. 'ml' or 'parts')
- Methanol: 23 (e. g. 'ml' or 'parts') and finally
- Water: 3 (e. g. 'ml' or 'parts').
The percentage content of your substrate we do not know (but me must not know that).
Imagine: if you want/need/are requested to make e. g. a 5% (w / vol or vol/vol) 'monophosphoryl lipid' reagent you take 5 g (or 5 ml, if the substrate is fluid) of your substrate to dissolve and FILL UP to 100 ml with your "solvent solution".
If you need then a dilution of 1:1 (if you have to dilute 5% to 2.5%) you just have to add the same amount of (the whole, combined) solvent to the "first = 100 ml of your 5% solution (=of your initial reagent + 100 ml of additional fresh 74:23:3 solvent).
Other dilutions you can calculate either bei cross-multiplication', 'rule of proportion', simply apply "cross-rule of mixture-operation" (x, y) , or automated calculation programs one can find in the www ....
Hope this explanation may loosen your short-term mental block or your blockade of thinking... (:-)) .... anyway: good luck and best wishes....
Thanks doctor for your helpful answer. I searched and decided to evaporate the solvent using a vaccum concentrator in order to use another more biologically compatible solvent the DMSO. However now i need to sterilize the resulting powder as i am going to use it in cell culture studies. Can you recommend a sterilization method?
Is it better to sterilize the powder or dissolute it firt and sterilize the liquid?
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