Hi,
I'm working on the immune cells in the heart and have a problem with CCR2 marker.
I did some research and found there's CCR2+ monocytes in heart failure mouse model. When I analysed CCR2 cells gated from Ly6chi monocytes, I couldn't find any CCR2+ population in all of samples, although I got lots of monocytes.
I'm wondering if it's true or I did something wrong.
Many thanks.
Hi Tingting,
For many chemokine receptors, you'd have to stain them at 37C (not at 4C or on ice). I successfully stained freshly isolated lymph node cells for CCR6 in HBSS buffer (calcium, magnesium, phenol red) in a 37C water bath for 1 hr. I assume it would work the same for CCR2. I hope this is helpful.
Zhixin
Hello Zhixin Jing
I didn't think about the temperature before because I also had to stain other markers and those worked well except CCR2. Did you stain all markers in 37C?
I stained for CCR7 only in 37C and wash and stain for all other markers in 4C.
Zhixin Jing I just found a nice paper which talked about the staining of chemokine receptors.
https://www.rndsystems.com/blog/heat-detecting-chemokine-receptors-live-cells
I guess maybe you also worked on CXCR4 (10.1016/j.celrep.2021.108733)? Did you also intubate in 37C?
Tingting Wang Thank you for sharing this.
I have always stained CXCR4 at 4C and have no problem, but have never compared 4C and 37C head to head.
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