Author Topic: Storage of CyDyes at -80'C  (Read 4174 times)

Stan.K

  • Newbie
  • *
  • Posts: 3
Storage of CyDyes at -80'C
« on: April 03, 2013, 12:19:11 PM »
I have not found this topic being discussed on this forum, so here is my question. Does anybody have experience with long-term storage of CyDyes in DMF at -80'C, at a working solution concentration of 0.4 mM? I will be using minimal labeling dyes from the trial kit, so I will be making the working solution right away, without making a stock solution first. I won't use all of the dye and I want to store the remaining unused dyes. I understand that the standard protocols call for the immediate use of the working solutions of CyDyes. There is no technical data that GE could provide me that would indicate that CyDye solutions in DMF cannot be stored for example for one-two weeks (maybe a month) at a working solution concentration of 0.4 mM, if stored at -80'C. I understand the issues with the stability of the fluorochrome, sensitivity of the reactive groups. The melting point of DMF is -60'C, so it will be frozen solid at -80'C, and as for NHS, I personally stored NHS in solution for years at -80'C without any loss of reactivity, and used it for amine coupling in biosensors.
My thinking is that the long-term (or at least relatively long-term) storage of 0.4 mM solution of minimal labeling CyDyes in DMF should be possible at -80'C, but I would like to find out if anybody has any experience with that. Any input would be appreciated.

dfried

  • Full Member
  • ***
  • Posts: 154
Re: Storage of CyDyes at -80'C
« Reply #1 on: April 03, 2013, 08:05:13 PM »
The issue is the presence of water, which will break down the dyes.  So also consider that with every freeze-thaw, you'll be condensing water out of the air (I suppose you can take measures to overlay with nitrogen or argon first).  I have always stored in the 10x concentration at -20 C and diluted only what was needed for the experiment, in this way they were stable for over a month at 10x at -20 C.  But I have no experience storing the 1x diluted dyes. For what it's worth, under standard operating procedures, I found that the Cydye signal was very comparable to Sypro Ruby counterstaining, so if the dyes started going off, you'd see more things in Ruby than with the Cydyes.  And if there's a problem, you'll likely see it with Cy2 first.

Hope that helps,
David


Stan.K

  • Newbie
  • *
  • Posts: 3
Re: Storage of CyDyes at -80'C
« Reply #2 on: April 04, 2013, 06:12:46 AM »
The issue is the presence of water, which will break down the dyes.  So also consider that with every freeze-thaw, you'll be condensing water out of the air (I suppose you can take measures to overlay with nitrogen or argon first).  I have always stored in the 10x concentration at -20 C and diluted only what was needed for the experiment, in this way they were stable for over a month at 10x at -20 C.  But I have no experience storing the 1x diluted dyes. For what it's worth, under standard operating procedures, I found that the Cydye signal was very comparable to Sypro Ruby counterstaining, so if the dyes started going off, you'd see more things in Ruby than with the Cydyes.  And if there's a problem, you'll likely see it with Cy2 first.

Hope that helps,
David



David,

Thank you for your input! Since I would be using anhydrous DMF to make the 0.4 mM working solution of the CyDyes, the only source of water would be the air, and I only plan to use the remaining dye once, so there would be only one freeze-thaw cycle. Since I will be using the trial minimal labeling dye kit (not the regular size kit), I only can make the working solution of 0.4 mM for use/storage. Of course another option would be to get a fresh kit for additional staining, but the issue is the cost of those kits. So the question would be how the storage of a concentrated stock solution in DMF at -20'C compares to the storage of a 1x solution in DMF at -80'C for the same time period of two weeks-one month regarding the effect on the stability and reactivity of the dyes. Looks like I would have to just try and see how it will work out for me, unless someone has actually tried storing the 0.4 mM working CyDye solution at -80'C and the level of the dye degradation proved to be unacceptable.

Stan.K

  • Newbie
  • *
  • Posts: 3
Re: Storage of CyDyes at -80'C
« Reply #3 on: May 06, 2013, 12:33:50 PM »
An update for anyone interested in the answer to my original question. I stored all 3 CyDyes at a working solution (1x) concentration in fresh anhydrous DMF at -80'C (DMF freezes solid at that temperature) in 1 ul aliquotes for 2 weeks and then used them. The dyes worked as they should, and at this point I don't see why they wouldn't work after being in storage in those conditions for a month. Since I stored them in 1 ul aliquotes, I used up the entire aliquote and did not re-freeze the dyes. I will post an update if and when I use the remaining aliquotes.

Allison Mackay

  • Newbie
  • *
  • Posts: 41
Re: Storage of CyDyes at -80'C
« Reply #4 on: July 24, 2013, 06:46:15 AM »
Our lab has experienced no problems with Cy dyes stored at working concentration at -80 for much longer than the directions recommend (ie >3 months).  I would not mix fresh dyes with older dyes in the same experiment, obviously.

Brixacier

  • Newbie
  • *
  • Posts: 4
    • Buy Replica Christian Louboutin Alti Gold Spikes 160mm Cork Pumps Natural Gold

Shoes CHRISTIAN LOUBOUTIN YOLANDA 120MM PATENT LEATHER PEEP TOE PUMPS BLACK, price $138.00.
If you  liked CHRISTIAN LOUBOUTIN YOLANDA 120MM PATENT LEATHER PEEP TOE PUMPS BLACK, click the "I want to buy" below to find out where it is most advantageous to buy shoes brand Fake Christian Louboutin



 
 
Category: Christian Louboutin Peep Toe
Brand: Best Fake Christian Louboutin

Tags product: CHRISTIAN LOUBOUTIN YOLANDA 120MM PATENT LEATHER PEEP TOE PUMPS BLACK, Christian Louboutin, Christian Louboutin Peep Toe, replica christian louboutin, christian louboutin replica, christian louboutin outlet, christian louboutin sale, christian louboutin shoes outlet, replica christian louboutin shoes, cheap louboutin outlet, christian louboutin fake, christian louboutin shoes sale, cheap christian louboutin shoes sale, cheap louboutin sale, christian louboutin knockoff, christian louboutin, louboutin shoes outlet, christian louboutin outlet online, louboutin, louboutin or, christianlouboutin, shoes christian louboutin, shoes by christian louboutin, shoes from christian louboutin, where can i get christian louboutin shoes, outlet louboutin, christian louboutin red shoes, red christian louboutin shoes, fake christian louboutins, christian louboutins fake, red louboutin
 
Hello users of life-sciences-forums.com.  I would like to introduce you to our new web shop, cheap replica brands direct from China.
Purchase goods direct from China - offering multiple products, also offering delivery services purchased anywhere in the world.  For retail customers  - it is an opportunity to buy cheap goods from China, we have created a catalog of extensive directory of links to our stores on this page.  If you are interested in wholesale supply of Chinese products, the managers clououtlet.biz can assist with finding a suitable product, direct from the factory and ordered to your doorstep.
 
An important factor is that - it's not just the site of goods from china, the company is headquarted in Guangzhou.  Wholesale clients have a unique opportunity to go directly to the producers, without the middle men involved.
Own office in china, this ensures you the security of the transactions, and our ensures applied to borth wholesale and retail customers.
 
Sometimes, potential buyers do not know where or how to find a product that meets one or another specific needs.  In such situations, you can contact our experts and we shall find the product which you are looking for.
 
We will find you any pair of shoes of any brand!
 
On this page we will provide one of the samples of our products
 
Our web-sites:
-louboutin shoes outlet
-christian louboutins fake
-red christian louboutin shoes
-where can i get christian louboutin shoes
Market China:
 
shoes louboutin
Promo-code: #jhiiiiljkh-99999-hndfdf