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No-wash fixation/permeabilization kit for RBC


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#1 Gregory Idelson

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Posted 20 December 2016 - 06:48 AM

Is there any no-wash kit that can be used for staining intracellular antigens in RBC? I need to fix the cells, permeabilize them and stain the intracellulat antigen, without washing. Beckman-Coulter provides something PerFix-nc kit for nucleated cells, but their kit includes the lysis of RBC. 

 

Thank you



#2 bob1

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Posted 20 December 2016 - 07:39 AM

The nucleated cells will be the usual immunology targets of what are generally known as "white blood cells" - the easiest way to get pure preps of these is to lyse the RBCs. 

 

How much of no washing are you meaning? Do you want to get rid of the serum and most of the WBCs? You can just wash the blood in PBS a few times to do this. You should then be able to fix etc - you don't need a kit to do this, just some buffers (PBS or TBS usually), fixative (formaldehyde, glutaraldehyde, methanol, etc.) permabilization solution (detergents usually in PBS or TBS but antigen dependent) and your antibodies.






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