ACK buffer

[svenson]

Hey,

I am an almost beginner in the field of Immunlogy and try to get rid of red blood cells by cell lysis after suspending whole spleens. The buffer I use for this purpose is a standard ACK buffer and I incubated previously for 5-10min on ice. Unfortunately, the only thing I get out of it is a pellet of debris.

1. is the incubation time really too long?

2. is it possible that the buffer decays by sitting on my lab bench for a few months?

 

Thanks

[dust mate]

Hey,

I am an almost beginner in the field of Immunlogy and try to get rid of red blood cells by cell lysis after suspending whole spleens. The buffer I use for this purpose is a standard ACK buffer and I incubated previously for 5-10min on ice. Unfortunately, the only thing I get out of it is a pellet of debris.

1. is the incubation time really too long?

2. is it possible that the buffer decays by sitting on my lab bench for a few months?

 

Thanks

 

prepare single cell suspension first and then use no more than 2-3ml buffer per spleen. Don't lyse more than 1 minute! This time is more than enough