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Annexin binding buffer (urgent advice)

concentration preparation

2 replies to this topic

#1 madelingirly

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Posted 18 January 2012 - 09:10 PM

Dear Guys,
I am new in flow cytometry and now one in my lab has any experience in it too,
I want to prepare annexin binding buffer, I found its concentration, but what I cant find are these concentration per not.
Could any one verify this meaning for me please.
I have attached the protocol of my Annexin-V fluos the reagent I have and it is written that I should have a binding buffer :
10mMole HEPEs.
140mMole NaCl, and 5mMole CaCl2.
my question these quantities per liter or what????
Sorry for silly question, but it made me confused.
Thank you in advance
Shimaa

#2 keefec

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Posted 20 February 2012 - 12:04 AM

These quantities should be in litre.

#3 almost a doctor

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Posted 20 February 2012 - 02:03 AM

View Postkeefec, on 20 February 2012 - 12:04 AM, said:

These quantities should be in litre.


That's not strictly true.

I assume when you say mMole you actually mean mMolar (or mM). In this case the final volume is irrelevant, whether is 1ml or 1litre of 10mM Hepes, it will still be 10mM. You do of course need to know the volume to prepare the reagents, but what will change is the amount of reagent (in grams) you need, the concentration should be the same. So basically, choose your required volume and calculate from there.

Now, if you are talking about mmoles is a different story, but I doubt is the case as buffers are always referred to in Molar concentration.

Edited by almost a doctor, 20 February 2012 - 02:03 AM.






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