Melanin extraction from melanoma cells, How? - (Jun/03/2008 )
Hi, I refer to the methods to extract melanin from B16 melanoma cell.
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
-sasoriza-
QUOTE (sasoriza @ Jun 3 2008, 05:26 PM)
Hi, I refer to the methods to extract melanin from B16 melanoma cell.
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
melanins are lypophilic and must be extracted from cell and organelle membranes; NaOH destructs membranes and proteins, and DMSO will, among others, solve melanin;
why are you worried about cell toxicity? do you like to extract melanin w/o destroying the cells?
-The Bearer-
QUOTE (The Bearer @ Jun 5 2008, 11:48 AM)
QUOTE (sasoriza @ Jun 3 2008, 05:26 PM)
Hi, I refer to the methods to extract melanin from B16 melanoma cell.
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
melanins are lypophilic and must be extracted from cell and organelle membranes; NaOH destructs membranes and proteins, and DMSO will, among others, solve melanin;
why are you worried about cell toxicity? do you like to extract melanin w/o destroying the cells?
No. I want to destroy the cells and extract the melanin from the cells.
So incubating it at 80 degree celsius for 1 hr just to help lyse the cells and dissolve the melanin content? I can incubate either at lower temperature for a longer time or at high temperature for a shorter time as long as the melanin is completely extracted and dissoved?
-sasoriza-
QUOTE (The Bearer @ Jun 5 2008, 11:48 AM)
QUOTE (sasoriza @ Jun 3 2008, 05:26 PM)
Hi, I refer to the methods to extract melanin from B16 melanoma cell.
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
Some use only 1N NaOH and incubate at 80 degree celsius for 60 min. Some use 1N NaOH + 10% DMSO and incubate at 80 degree ceisius for 60 min.
May I know what is the purpose of adding DMSO? DMSO is toxic to the cells at high temparature?
melanins are lypophilic and must be extracted from cell and organelle membranes; NaOH destructs membranes and proteins, and DMSO will, among others, solve melanin;
why are you worried about cell toxicity? do you like to extract melanin w/o destroying the cells?
Can I use either 1N NaOH or 100% DMSO to dissolve the melanin contents?
-sasoriza-