Serum-plasma dilution in ELISA sandwich - (Feb/07/2007 )
I am setting up an ELISA sandwich to determine plasma-serum levels of a protein which (I suspect) is present at very small concentration. On one hand I want to dilute serum as least as possible to avoid the risk of being below the sensitivity level. But on the other hand I suspect that a not enough dilution of serum can result in unspecific and unacceptable background due specially to albumin???
Any idea from people experienced in dealing with ELISA sandwich using sera?
Thanks indeed in advance
-rammondia-
QUOTE (rammondia @ Feb 7 2007, 01:18 AM)
I am setting up an ELISA sandwich to determine plasma-serum levels of a protein which (I suspect) is present at very small concentration. On one hand I want to dilute serum as least as possible to avoid the risk of being below the sensitivity level. But on the other hand I suspect that a not enough dilution of serum can result in unspecific and unacceptable background due specially to albumin???
Any idea from people experienced in dealing with ELISA sandwich using sera?
Thanks indeed in advance
Any idea from people experienced in dealing with ELISA sandwich using sera?
Thanks indeed in advance
I usually do a serial dilution starting off 1:1 and then 1:5 and 1:25 that way you can dilute out the sera and hopefully catch your protein of interest. It has worked nicely for me in the past.
-tlblase-