Hi there..this is my first time to handle reagents..we just bought Hydrocortisone and EGF from Sigma for our cell culture project
problem is i can't figured out how to reconstitute them, the vial doesnt come with any manual, sigma webpage only said to add absolute ethanol (Hydrocortisone) and acetic acid (EGF)
somebody has a tutorial? my working solution should be 10 ug/ml (Hydrocort) and 10 ng/ml (EGF)
doesn't ethanol and acetic acid harmful for cell?
thanks a lot !
let say i have 10 mg of powdered hidrocort, sigma instruct to reconstitute in EtOH and dilute in sterile medium..
if i add 10 ml of EtOH and dilute with 10 ml of Sterile culture medium to achieve 500 ug/ml stock solution, aliquote and stored in - 20 C ? is this procedure alright?
other intruct to add 0,1 % BSA...means?
check for any published paper for their solubility and stock concentration.
ethanol does effect the vialibity in cell culture. so better work on this thoroughly and make a stock
just wondering if i aliquote and freeze reconstitute HC in EtOh, without further dilution using medium, will it be ok ? i plan to dilute it later after thawing..
Hi! I have bought the same products I have the same problem, I don't know if etanol or acetic acid is harmful to cells and I dont know exactly how to reconstitute them, because the concentration needed is to low, and the stock solution proposed is too high to only dilute it in the medium. If someone could inform me and give some light in this i would apppreciate it. is the fiorst time I used these compounds. I dont understand very well the data sheet information provided by sigma.
Thanks!
Hi Ana, the question really brings back memory :) almost a decade since I start working in lab
I already forgot the solution used spesific for this question, but I do know now from experience that EtOH is cytotoxic and you best keep it under 1 % of medium
Acetic acid change medium pH so you also want to keep it minimal, best is you reconstitute with EtOH and dilute further with medium to your working concentration
Good luck to you
Thank you very much for your answer!! I will dilute it at maximun for not harm cells
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