It's is well known erythrocytes have got large amount of glutathione (GSH) and particular condition,such as oxidative stress, can deplete GSH changing in its disulfide form GSSG.For this reason, it is well recommended an accurate techniques like flow cytometry in order to determine the level of GSH.
Dear Arbace,
you better avoid Mercury orange method, because it requires immense washing and cell will lost. Better if you stain with CMFDA and measure by 488 Argon laser in flow cytometry, it is easier and better method. If you want protocol, please let me know.
BW,
Debanjan
Dear Debanjan,
thanks for your response.I have never used this reagent but as you told me,I will try.
Please if you can send me protocol.Anyway, if I have questions I will contact you,sure.
My mail si [email protected]
Dear Arbace,
I will send you the protocol for the CMFDA staining through email.
Best,
Debanjan
I agree with Debanjan to avoid Mercury orange also because it is less consistent and has higher background.If you want to use CMFDA method you should aware this method reflects the total level of free intracellular thiol rather than GSH specifically you might use it with sample of cells that have been depleted of GSH or compare it with other staining methods such as Monobromobimane
Dear Arbace,
I agree with Dr. Hunaiti too. I sent you both the protocol for Monobromobimane and CMFDA. Please feel free to discuss.
I´m also interesten in your protocols. I´ve tried, but i´m mot so happy about standardisation
Hi Debanjan Mukhopadhyay
I'm also interested by your protocol for Monobromobimane.
Is it possible for you to send it at : [email protected]
Thank you
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