# 5' aza-cytidine - stock solution - (Feb/01/2010 )

Hi all,

I'm gonna start some dnmt inhibitor experiments. Can someone tell me how to make a stock solution ? how stable it is ?? i have 10 mg solid 5 aza2dc from sigma chemicals. thanks in advance.

RKA on Feb 1 2010, 07:07 AM said:

Hello!

For our 5Aza, we use the following as a stock:

__100,000X Stock__-Dissolve 1 mg of 5-Aza-2dc in 4.380 mL of STO* media (w/o FBS)

-Aliquot in sterile microfuge tubes, 50µl per tube

-Store in -80

no idea how stable, but at -80 I'd say at least a year

Then for the working solution we use:

__Working Solution (1X=0.01µM)__-Make a 1:10 dilution (10,000 X) by taking 10µl of 100,000X stock and adding 90µl of STO media

-Make another 1:10 dilution (1,000X) by taking 10µl of 10,000x stock and adding 90 µl of STO media

-Make another 1:10 dilution (100 X) by taking 10µl of 1,000x stock and adding 90 µl of STO media

-Finally, make another 1:

__100__dilution by taking 10µl of 100x stock and adding 990 µl of STO media

*STO is what we use as a cell culture media.

-100mL Glasgows MEM

-1.2mL of 200mM L-Glutamine

-1.2mL of 100mM Sodium Pyruvate

-1.1mL DMEM

thanks for the answer, everyone using different solvent to dissolve 5AZA, like ethanol, dmso etc...doo u think that doesent matter ??

k_undertoe on Feb 1 2010, 07:07 PM said:

RKA on Feb 1 2010, 07:07 AM said:

Hello!

For our 5Aza, we use the following as a stock:

__100,000X Stock__-Dissolve 1 mg of 5-Aza-2dc in 4.380 mL of STO* media (w/o FBS)

-Aliquot in sterile microfuge tubes, 50µl per tube

-Store in -80

no idea how stable, but at -80 I'd say at least a year

Then for the working solution we use:

__Working Solution (1X=0.01µM)__-Make a 1:10 dilution (10,000 X) by taking 10µl of 100,000X stock and adding 90µl of STO media

-Make another 1:10 dilution (1,000X) by taking 10µl of 10,000x stock and adding 90 µl of STO media

-Make another 1:10 dilution (100 X) by taking 10µl of 1,000x stock and adding 90 µl of STO media

-Finally, make another 1:

__100__dilution by taking 10µl of 100x stock and adding 990 µl of STO media

*STO is what we use as a cell culture media.

-100mL Glasgows MEM

-1.2mL of 200mM L-Glutamine

-1.2mL of 100mM Sodium Pyruvate

-1.1mL DMEM

i don't think it matters. i dilute it in DMSO because my vehicle contains DMSO as well. also, i thought i saw on the sigma site that it's only stable for a month at -80.

RKA on Feb 2 2010, 02:43 AM said:

k_undertoe on Feb 1 2010, 07:07 PM said:

RKA on Feb 1 2010, 07:07 AM said:

Hello!

For our 5Aza, we use the following as a stock:

__100,000X Stock__-Dissolve 1 mg of 5-Aza-2dc in 4.380 mL of STO* media (w/o FBS)

-Aliquot in sterile microfuge tubes, 50µl per tube

-Store in -80

no idea how stable, but at -80 I'd say at least a year

Then for the working solution we use:

__Working Solution (1X=0.01µM)__-Make a 1:10 dilution (10,000 X) by taking 10µl of 100,000X stock and adding 90µl of STO media

-Make another 1:10 dilution (1,000X) by taking 10µl of 10,000x stock and adding 90 µl of STO media

-Make another 1:10 dilution (100 X) by taking 10µl of 1,000x stock and adding 90 µl of STO media

-Finally, make another 1:

__100__dilution by taking 10µl of 100x stock and adding 990 µl of STO media

*STO is what we use as a cell culture media.

-100mL Glasgows MEM

-1.2mL of 200mM L-Glutamine

-1.2mL of 100mM Sodium Pyruvate

-1.1mL DMEM

Hi,

I am currently using Sigma 5-AZA that was diluted as described in the post above in PBS in april 2009. It still works fine...

Tobi