Low ELISA titers - (Nov/18/2005 )
Hi,
I have many antisera projects that are coming up with lower ELISA titers than are expected. I realize that there are several factors that could cause this, such as antigenicity of the peptide, conjugation. However, I was wondering what others would consider a usable titer reading? I'm satisfied when my titer readings are 1:100,000 and higher, but recently I have found them to be 1:4000 to about 1:20,000. Are these usable for affinity purification purposes? Will I obtain any purified AB? I would like to know about the experiences of others since I am fairly new to immunology.
Any input would be greatly appreciated.
Thanks,
Abby
-abby1-
QUOTE (abby1 @ Nov 18 2005, 06:45 PM)
Hi,
I have many antisera projects that are coming up with lower ELISA titers than are expected. I realize that there are several factors that could cause this, such as antigenicity of the peptide, conjugation. However, I was wondering what others would consider a usable titer reading? I'm satisfied when my titer readings are 1:100,000 and higher, but recently I have found them to be 1:4000 to about 1:20,000. Are these usable for affinity purification purposes? Will I obtain any purified AB? I would like to know about the experiences of others since I am fairly new to immunology.
Any input would be greatly appreciated.
Thanks,
Abby
I have many antisera projects that are coming up with lower ELISA titers than are expected. I realize that there are several factors that could cause this, such as antigenicity of the peptide, conjugation. However, I was wondering what others would consider a usable titer reading? I'm satisfied when my titer readings are 1:100,000 and higher, but recently I have found them to be 1:4000 to about 1:20,000. Are these usable for affinity purification purposes? Will I obtain any purified AB? I would like to know about the experiences of others since I am fairly new to immunology.
Any input would be greatly appreciated.
Thanks,
Abby
I think you will be fine. You will obtain purified Ab with lower concentration.
After loading onto the affinity column, you may wish to check your flowthrough with ELIZA to see whether there is still some Ab. If there is still a lot of Ab, reload the column.
I hope this may help.
-Minnie Mouse-