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!
Photo
- - - - -

Seeking advice on coated plates storage

coating storage

  • Please log in to reply
5 replies to this topic

#1 RILC

RILC

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 27 April 2012 - 03:17 PM

Hi. What are recommended (from your experience) storage conditions for coated ELISA plates? I coat them overnight, in the fridge, with BSA-hapten in 50 mM TRIS pH 9.
Can they just be washed (I use low strength TRIS), sealed and put in -20°C freezer?
Should they better be blocked (I use 1% casein in TRIS) and washed before storage?
Is it better to dry (method?) them?
I'd love to hear your opinions.

#2 Ben Lomond

Ben Lomond

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 75 posts
6
Neutral

Posted 29 April 2012 - 10:12 AM

there was a thread from November last year with some advice

#3 PAO_ahac

PAO_ahac

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 94 posts
0
Neutral

Posted 30 April 2012 - 03:50 AM

Coat plates the way you normally would. Then block. After decanting fill with your buffer containing 5% sucrose in your blocking buffer; allow to sit 30-1 hr rt. then decant. do not wash do not rinse. dry inverted overnight. Store in sealed plastic bag with dessicant pouch at RT or 4C. This is equivalent to what manufacturers do to produce kits.

You may have to optimize some of these steps since it has been quite some time since i did this. Crucial component is the sugar and bsa as protectants.

#4 Ben Lomond

Ben Lomond

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 75 posts
6
Neutral

Posted 30 April 2012 - 02:55 PM

With that said....it is my experience that precoated and dried plates rarely match up to those that have been coated the day (or a few days) before and left in the coating buffer. On a number of occasions, I have asked reputable kit manufacturers to keep their pre-coated plates and provide me with the coating antibody in solution resulting in reduced CVs between wells.

#5 PAO_ahac

PAO_ahac

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 94 posts
0
Neutral

Posted 01 May 2012 - 04:58 AM

If your use is R&D only then leaving the wet solution in sealed plates will be fine. However, if you are storing for weeks, months, to 1 yr then dry is the best approach. Commercial, clinical lab kits with dried plates provide documentation of CV's less than 5%. Additional specifications are usually detailed in kit protocols that are approved by FDA 510(k) process.

#6 RILC

RILC

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 01 May 2012 - 04:31 PM

Thanks a lot for your advice/suggestions! I didn't want to use preservatives (we have thimerosal and sodium azide, the latter wouldn't be that suitable since I use HRP, though a good washing technique should remove all residues) so I had thought of freezig or drying. I'll see if I can test the different aproaches.





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

©1999-2013 Protocol Online, All rights reserved.