I have whole blood and want to isolate rbc. I've come across some methods that isolate wbc, but haven't been able to find one for the collection of rbc. Any help would be appreciated.
See paper:
Nutrients. 2014 Mar 20;6(3):1164-80. doi: 10.3390/nu6031164.
Dietary zinc deficiency affects blood linoleic acid: dihomo-γ-linolenic acid (LA:DGLA) ratio; a sensitive physiological marker of zinc status in vivo (Gallus gallus).
Reed S1, Qin X2, Ran-Ressler R3, Brenna JT4, Glahn RP5, Tako E6
It's not my project, I'm just helping someone find a protocol, but essentially, she wants to see if there's any difference in qPCR signal in her samples when removing (or keeping) white blood cells. We've come across methods where they spin whole blood in leucosep tubes, and it isolates wbc, but what she wants, the rbc, are below the barrier.
You can have a look at this paper, it describes a simple and reliable method for isolating erythrocytes:
Beutler E, West C, Blume KG. The removal of leukocytes and platelets from whole blood. J Lab Clin Med 1976; 88: 328–33.
Standard protocol of RBC separation (based on centrifugation 500-750g X 5-10 min ) from whole blood included:
1. Separation of RBC from plasma;
2.Washing with buffer (PBS or HEPES or ....) two-three time.
Available from: https://www.researchgate.net/post/Is_there_a_specifc_standard_method_to_separate_erythrocytes_from_whole_blood_samples_for_further_analysis [accessed Apr 27, 2015].
Leukodepletion and platelet removal would be useful for you, right? Attached are two protocols. One is for the RBC filtration through the cellulose column (self-made), the other is for more advanced separation of RBC frations (low, medium and high denisty) on Percoll density gradient. If you want to be completely WBC-free, go for filtration followed by separation of the gradient. UZH is University of Zurich :)
More information on the issue is within the open-access review to which we have also contributed: http://www.sciencedirect.com/science/article/pii/S0268960X13000064
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