Important Notice: Our web hosting provider recently started charging us for additional visits, which was unexpected. In response, we're seeking donations. Depending on the situation, we may explore different monetization options for our Community and Expert Contributors. It's crucial to provide more returns for their expertise and offer more Expert Validated Answers or AI Validated Answers. Learn more about our hosting issue here.

What is the best way to maintain cell-surface staining with Perm Buffer III?

April 26, 2017best cell-surface maintain staining

0

Posted

What is the best way to maintain cell-surface staining with Perm Buffer III?

1 Answer

0

Posted

Many times the regular fixation/permeabilization protocol with Perm Buffer III (Cat. No. 558050) completely obliterates surface staining for some antigens such as human CD16, CD19, CD56, and CD14. Validated Cell Surface Markers To maintain cell surface staining, investigators can activate cells to induce phosphorylation, fix the cells, stain for surface antigens, then permeabilize the cells with Perm Buffer III. Surface staining will decrease in intensity (separation between negative and positive population will decrease) but remains detectable. This modified protocol has been tested in house with anti-human CD16, CD14, CD19, and CD56.