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Help with the Red Blood Cell Lysis

Flow cytometry Monoclonal antibodies Inmunophenotyping Buffer Solutions Lysing solutions

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#1 ClaudiaSul

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Posted 17 September 2016 - 09:59 AM

Hello I am writing from Peru. Currently I am working in a Flow cytometry laboratory in oncohematology. The question is that we work with the Euroflow consensus and of course we work with manufacturers, clones, quantities includes in its protocol, the question is that we want to change the manufacturer (the mayority of reagents and solutions are BD) in our panels, so we start with some antibodies and we noticed that in some other manufacturers, using the same clone, the same quantity, and the same fluorescence we found that the MFI in some antibodies are dicreased in comparison with the clones and marks that we actually use in laboratory. Another issue is in the use of red blood cell lysing, we found that some subsets are lower than when we use the actual lysing. And a finally question, is incorrect to say " that reagent doesn't work" because all manufacturers in the world invest to produce and sell their product, that of course have to work. But why are that kind of differences, the method doesn't reproduce in my lab, althought I follow their protocol and Technical data sheet step by step.

 

I hope someone can help me. I am really desperate.

 

Thanks a lot

 

Claudia Sul



#2 bob1

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Posted 19 September 2016 - 07:24 AM

The answer to this is that not all antibodies are equal, and it is also the reason why some antibodies from individual suppliers are certified for use in clinical labs but the same antibody from another is not. Those antibodies that are certified are checked batch to batch to ensure consistent signal, whereas non-certified will depend on the animal and processes used for preparation. In non-clinical settings, this variation doesn't matter all that much, but it is all important for clinical data. If you are working in a clinic, you should use the certified ones, they are more expensive, but they are guaranteed to work.

 

Manufacturers do invest time and money in making sure that their things work, so if it fails in your lab, it may be that your lab is just not doing something correctly, or that there is something small that you've missed. Having said that, lab to lab reproducibility is a problem for many things, so it may be that there is some process in your lab that isn't compatible with the reagents.







Also tagged with one or more of these keywords: Flow cytometry, Monoclonal antibodies, Inmunophenotyping, Buffer Solutions, Lysing solutions

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