Hi, I am preparing media for cell culture (murine immune cells) and I wanted to know what is the maximum amount of 2-mercaptoethanol in a 500 mL media cell culture stock. Usually I would add 50 microliter/50mL of cell culture media but can I add more since I read it enhances antigen presentation. Note that the stock of 2-ME I am using is at 1000x.
Ty
2-ME is also very toxic, I did not calculate the amount you added, but it seems that you add too much.
Usually, I prepare a 2ME stock solution from original (pure, 14.3 M concentration) at 1 to 100. Then add 200 ul of the stock solution to 500 ml medium.
Good luck
We are using 50 µM 2-ME for murine B-cells.
Xianzhu Wu's dilution also results in 50 µM final concentration.
This review should be helpful.
Article Review: 2-Mercaptoethanol alteration of in vitro immune func...
Xianzhu Wu , we are also using final concentration of 50 µM for T cells and dendritic cells.
That concentration should be fine. Only thing is that the stock solution should not be kept too long, I usually prepare every 3 months.
According to your desription, you have added 1 microl/ml (dilution 1:1000 from the concentrate commercial solution). So, 14.3 M is diluted to 14.3 mM. This is a lot, probably toxic for cells (I suppose that this is depending of cell type and purpose). Normal additions to cell culture (melanocytes when I used) is between 0.05 and 0.1 mM. Cheers
Hi, Francisco Solano
I never added 1 ul /ml. What I described was " original (pure, 14.3 M concentration) at 1 to 100. Then add 200 ul of the stock solution to 500 ml medium. " That's 50 uM. Actually I tested the concentrations to culture macrophage using concentration between 25 to 100 uM (i.e 0.025 to 0.1mM), 50 uM still the best although 25 and 100 uM still works.
14.3 mM concentration will kill all cells.
Ok we agree. I misunderstood the sentence " Usually I would add 50 microliter/50mL of cell culture media. This is the same than 1 microliter/ml. The point is the previous dilution.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology
- Enzymes