Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

Anyone familiar with ELISA of cytokines in mouse plasma?

  • Please log in to reply
1 reply to this topic

#1 Nrelo



  • Active Members
  • PipPipPipPipPip
  • 71 posts

Posted 19 January 2012 - 04:48 AM

I am new to ELISA and I am going to measure TNF-alpha in mouse plasma (from heparinized blood).

The blood was collected from terminal bleeding and I got about 300~400ul plasma. I am not sure about what diluent I should use for diluting the samples.

Also, does anyone know about the basal level of TNF-alpha (in pg/ml) in mouse plasma when doing ELISA?

Many thanks!

#2 Martin Fitzpatrick

Martin Fitzpatrick


  • Active Members
  • Pip
  • 12 posts

Posted 19 January 2012 - 05:40 AM

One option if you are unsure of the concentration is to start out plating samples twice - one at the neat concentration, one at 1 in 10 dilution (10ul + 90ul) - using the same standard. If the concentration is particularly high the neat plate will develop very quickly, but the 1/10 dilution should remain within the standard curve. If you're really unsure you can do further dilutions 1/100, 1/1000 etc. Do this on a small scale so as to not waste your sample.

For diluting you can use the same solution you use to dilute the standards (commonly called Assay Diluent in kits) or normal cell media (RPMI) have both worked fine for me.

When it comes to analysis I've got a plate calculator available here that allows you to put different plate dilutions in.

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.