Hi Saquib !!

I also work with ascorbic acid in my lab, and yes... you just have to watch it and it get oxidized ! In my experience, the best you can do is to prepare your standard solution of ascorbic acid in an amber tube, or in case you have to prepare a large amount, you can put some foil around a falcon tube, to protect your preparation from the light. Then, put it on ice (buried, so it also protects from light) and I personally put it inside a drawer or a dark place until I use it.

Since I don't have to prepare a very concentrate standard of ascorbic acid (I just weight a few mg), I prepare a new standard everytime I have to do an experiment. I don't put it in dark and store it at 4°C or -20°C, because I don't want to take the risk of using a solution that is not 100% ascorbic acid.

If you are using cells and you have to pre-treat them with a solution of ascorbic acid, just prepare your standard, then make the dilution and then put it into your cells. Leave the cells in the incubator, they will have a good tempeture and also inside of it, it's dark.

I hope I have help you in some way :) And good luck !

Paulina

Hi Saquib !!

I also work with ascorbic acid in my lab, and yes... you just have to watch it and it get oxidized ! In my experience, the best you can do is to prepare your standard solution of ascorbic acid in an amber tube, or in case you have to prepare a large amount, you can put some foil around a falcon tube, to protect your preparation from the light. Then, put it on ice (buried, so it also protects from light) and I personally put it inside a drawer or a dark place until I use it.

Since I don't have to prepare a very concentrate standard of ascorbic acid (I just weight a few mg), I prepare a new standard everytime I have to do an experiment. I don't put it in dark and store it at 4°C or -20°C, because I don't want to take the risk of using a solution that is not 100% ascorbic acid.

If you are using cells and you have to pre-treat them with a solution of ascorbic acid, just prepare your standard, then make the dilution and then put it into your cells. Leave the cells in the incubator, they will have a good tempeture and also inside of it, it's dark.

I hope I have help you in some way :) And good luck !

Paulina

Dear paulina,

Thanks a lot for you valuable reply, can you tell me how many concentration you are using for cell treatment, and after how many time you are treating again i meany every after 24 hours or after 48 hours?

As you know if i will use 100-200 micro molar/ml then it is very low concentration and to make this solution is very difficult.

Please suggest

thanks

Dear Saquib:

I don't really know which kind of experiment are you doing, but I'm measuring the efflux of Ascorbic acid from Astrocytes under glutamate stimulation. I pre-treated my cells with 1 mM ascorbic acid in IB (incubation buffer) for 1 hour. I put the cells in the incubator, so they were at 37°C and in total darkness (so the ascorbic acid was not oxidized).

I suggest you look in some papers or ask somebody the concentration of ascorbic acid you need, depeding in the experiment you are doing.

Good luck and if you have more questions let me know :)

Paulina.

Are you preparing Ascorbic acid in water or something else???

Dear,

Prepare and use quickly dont store

Do not store it, it better to use it fresh each time.

Better if you can add it in a solid form if this OK with your experiment.

Dear Ashraf,

I want to treat my cells by Sodium L-Ascorbate. But for making solution which one is better, PBS or Water?

Thank in advance...

Dear Puspa

Usually I prepared the solution in MiliQ water

Dear Ashraf,

It sounds great to get a quick answer. I need to prepare 5 - 80 micromolar solution. But its so small amount and I need to prepare stock solution. So usually what you did for storing your stock solution and also do I need to pay more attention for my cells during incubation like covering or something like that?

Dear Puspa

Prepare 2-5 ml of stock solution don't store it because it will be degrade, use as per your requirement and discard the rest.

Try to add ascorbic acid in the last just before the incubation to avoid the light.

hi there I want to use ascorbic acid as a reductant, so what are the concentrations that you suggest??

Please, can someone help me with the standard concentration ascorbic acid and iron Sulphate need for in vitro malaria work??

Please, can someone help me with the standard concentration of ascorbic acid and iron Sulphate need for in vitro malaria work??