Advertisement
If you have a new account but are having problems posting or verifying your account, please email us on hello@boards.ie for help. Thanks :)
Hello all! Please ensure that you are posting a new thread or question in the appropriate forum. The Feedback forum is overwhelmed with questions that are having to be moved elsewhere. If you need help to verify your account contact hello@boards.ie

Buffers!

Options
  • 10-11-2008 11:17pm
    #1
    Galmay
    Registered Users Posts: 66 ✭✭


    Hi everyone,
    Im starting a MSc by thesis soon. I'll be carrying out cell separations using magnetic microbeads on an automated system. The company that manufactures the microbeads recommends a PBS buffer of pH 7.2 and 2mM EDTA both with and without 0.5% BSA.
    Conveniently, the company sells both of these pre-made, however at £90stg a pop for 3 1500ml bottles of each, it isnt cheap. The problem though is that the buffer containing the BSA is good for only 3 days once opened. Considering i will be carrying out maybe one separation every 2 weeks, there is potentially a lot of wastage.
    I was looking into just buying the buffer without BSA and buying in BSA stock soln and prepare it myself as needed, but the quotes Im getting for the stock solution is very expensive. As Im working in a routine hospital lab environment, pH meters, weigh scales, graduated glassware etc and any other basics for preparing my own buffers are thin on the ground, plus that is very time consuming and i have my day job to do also!
    So anybody out there who has experience in this area got any ideas for me to cut down on costs/waste? Or is this the best deal I can hope for? Any help at all is very much appreciated!


Comments

  • Options
    #2
    2Scoops
    Registered Users Posts: 1,845 ✭✭✭2Scoops


    BSA is expensive but gets cheaper if you get a less 'pure' product. It will also be cheaper if you buy it as a desiccated protein and not in solution. This might lead to problems if you don't have adequate facilities to prepare solutions, though.

    The good news is that if it's only being used as a buffer for a cell isolation procedure, being a little off on the concentrations won't be a big deal. You could probably* even do it in PBS with EDTA and no FBS, as long as it is a short separation time-wise.

    *Edit: Actually, don't quote me on that - depends on your endpoint.


  • Options
    #3
    GuanYin
    Closed Accounts Posts: 15,552 ✭✭✭✭GuanYin


    Can you freeze the buffer in aliquots?


  • Options
    #4
    sarumite
    Registered Users Posts: 2,909 ✭✭✭sarumite


    I have done separation with micro beads in the past. Personally I wouldn't buy ready made from a manufacture as it is always a total rip off....and their expiry dates are always really short.

    I would buy in some PBS (cell culture grade.....though I use tablets resuspended in 18 M-Ohm (milliq)water and then autoclaved). Then I would prepare a solution of EDTA and BSA and filter it through a .2 um syringe filter and add it to the PBS. That is how I would do it, however you are responsible for your own work, so best do it the way you feel happiest with


Advertisement