Serum or Plasma, which is better to detect cytokines? - additionally, how about the comparation of Luminex and ELISA? (Feb/10/2006 )
Hi friends,
I need to detect cytokines like IL1b, IL6, TNFa, etc. in human blood
I've got serum and plasma from patients. Which will be better for the purpose, serum or plasma?
How about the comparation of Luminex and ELISA?
Thanks for response.
yxz,
You just asked the most common question I get from customers regarding Human ELISA kits.
Sample handling is the name of the game. There are a handfull of papers out trying to correlate the conc's of cytokines in serum to plasma and determine if one is better for ELISAs. Some say there is no difference and other say there is a large difference. My experience is that 95% of time, the researcher uses the method they are most comfortable with or that which their colleagues in their field use most often. And, unsurprisingly, that often works just fine.
Regardless, the best way to limit problems (no matter which method you choose) is to immediately start separating the plasma or serum (post clotting) from the cellular elements. This provides optimal analyte stability and is the biggest red flag when I talk to researchers about their protocols.
Other than that follow your protcol as closely as possible. That will give you the most reproducible results and will either give you reproducible results or let you know you need to switch sample types.
To make a point: the ELISA kits you purchase (at least ours!) are put through rigorous testing with serum, plasma, etc. before being brought to market . Hope that gives you some confidence.
Best of luck.
cg
Product Manager, Diaclone
diaclone@tepnellifecodes.com
Thanks a lot for your detail reply. 
You just asked the most common question I get from customers regarding Human ELISA kits.
Sample handling is the name of the game. There are a handfull of papers out trying to correlate the conc's of cytokines in serum to plasma and determine if one is better for ELISAs. Some say there is no difference and other say there is a large difference. My experience is that 95% of time, the researcher uses the method they are most comfortable with or that which their colleagues in their field use most often. And, unsurprisingly, that often works just fine.
Regardless, the best way to limit problems (no matter which method you choose) is to immediately start separating the plasma or serum (post clotting) from the cellular elements. This provides optimal analyte stability and is the biggest red flag when I talk to researchers about their protocols.
Other than that follow your protcol as closely as possible. That will give you the most reproducible results and will either give you reproducible results or let you know you need to switch sample types.
To make a point: the ELISA kits you purchase (at least ours!) are put through rigorous testing with serum, plasma, etc. before being brought to market . Hope that gives you some confidence.
Best of luck.
cg
Product Manager, Diaclone
diaclone@tepnellifecodes.com
It depends. If you want to measure alot of cytokines in the same sample and have limited volume of the sample I'd say to use luminex, if you have access to a luminex machine. If you have more sample and want to measure only 1 or 2 cytokines it's probably more cost effective to use ELISA kits. You can buy kits with the abs and standard and coat your own plates which reduces the price.
All the best,
Ceri
All the best,
Ceri
Ceri is right: Multiplex ELISAs are great tools, particularly if you have limited sample volume. It's hard to get more than a few hundred microliters of serum from a mouse without sacking it, so getting serum concentrations of multiple cytokines out of a mouse is impossible without multiplexing. So, if you don't have Luminex flow cytometer, but you do have access to a gene microarray scanner (pretty much anything except Afffymetrix) or a Li-Cor Odyssey machine, you can use a slide-based array from RayBiotech. They have quantitative arrays called "Quantibody" that can measure 60 proteins at low pg/mL concentrations in less than 100 uL sample volume. I think they have testing services too.
Cheers,
tiger