BSA: Bovine serum albumin
I would like to grow my cells in DMEM without serum but with BSA.
There are two forms of protein supplement (e.g. BSA) in the market, one is lyophilised powder and one is liquid.
How should i add them into the DMEM?
for example, the BSA powder come in 1G. if i wanna make 1% w/v of BSA in DMEM, how should i make it? do i dilute first and then add into DMEM as a solution (do in 1% v/v?) or do i straightly add the powder into the DMEM?
if the BSA or other supplement protein come in liquid form, e.g. in 250 mg/mL (packaging of 1mg --> not sure what this means)
how should i add this protein solution into DMEM if i wanna make 1% v/v of BSA in DMEM? or do i need to calculate based on concentration (mg/mL or ug/mL)?
I am confused about the calculation as most of the published paper did not write it out clearly how did they do.
BSA is measured by weight by volume (powder form). For example, to make a 100mL 3% BSA solution, use 3 grams of BSA and bring the volume to 100mL. To make a 5% solution of BSA in 500mL of media (which is the typical media bottle size) 25grams of BSA would be used, brought to a volume of 500mL with the media of interest.
In the case of the 250mg/mL bottle of BSA that you have, you will need to figure out a specific concentration of BSA you would like in the media (using the C1V1=C2V2 equation)... Are concentrations published? You have a 25% stock solution of BSA. You can make a calculation based on this number. I found this protocol for media with a 25% stock solution: https://www.helsinki.fi/sites/default/files/atoms/files/reagents_and_media_preparation_.pdf
I hope this helps!
http://www.iun.edu/~cpanhd/C101webnotes/aqueoussolns/wvpcnt.html
BSA is measured by weight by volume (powder form). For example, to make a 100mL 3% BSA solution, use 3 grams of BSA and bring the volume to 100mL. To make a 5% solution of BSA in 500mL of media (which is the typical media bottle size) 25grams of BSA would be used, brought to a volume of 500mL with the media of interest.
In the case of the 250mg/mL bottle of BSA that you have, you will need to figure out a specific concentration of BSA you would like in the media (using the C1V1=C2V2 equation)... Are concentrations published? You have a 25% stock solution of BSA. You can make a calculation based on this number. I found this protocol for media with a 25% stock solution: https://www.helsinki.fi/sites/default/files/atoms/files/reagents_and_media_preparation_.pdf
I hope this helps!
http://www.iun.edu/~cpanhd/C101webnotes/aqueoussolns/wvpcnt.html
Hi dear,
I recomiended steril liquido solution and how Elliot recommended just you have to calculated the volume necesary of the BSA and complete with culture médium. All this process in the biosegurity cabine. At the end your medium is steril.
If you jus have BSA in power presentation, you have to calculate the weigth of BSA and to let to disolver in médium and then to complete the exact médium in a volumétric container. At the end you should to sterilized by filtration.
Bye
A 1% w/v solution is 1 gram/100 ml, so 1 gram BSA/100 ml DMEM. Purified BSA powder should easily dissolve in DMEM with gentle stirring. Be careful with filter sterilization afterward to not get a lot of foaming from the dissolved BSA. A 250 mg/ml stock solution of BSA is 25%; you would dilute it 1/25 (i.e. 40 ml/L) in DMEM to achieve a 1% BSA in DMEM. If you use a commercial BSA solution, be sure it does not contain sodium azide as a preservative. Sodium azide would poison your cells.
If you want 1% of BSA in DMEM,
i.e. weight/volume of BSA,
1g of BSA in 100ml DMEM.
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