hello
I am studying neutrophil and need specific marker for activated one ( nucleic or cytoplasmic markers) for FACS, IF applications. Thank you
Dear Iman Aldybiat,
You may look over the following info:
https://onlinelibrary.wiley.com/doi/10.1002/eji.201646632
What are flow cytometry markers?
These markers are usually functional membrane proteins involved in cell communication, adhesion, or metabolism. Immunophenotyping using flow cytometry has become the method of choice in identifying and sorting cells within complex populations, for example the analysis of immune cells in a blood sample.
https://www.abcam.com/protocols/flow-cytometry-immunophenotyping
What is the principle of FACS?
Flow cytometry (FCM) is a technique which enables rapid analysis of statistically significant number of cells at single cell level. The main principle of this technique is based on scattering of light and emission of fluorescence which occur when a laser beam hits the cells moving in a directed fluid stream. Feb 5, 2021
What are the protein markers of a cell?
Cell-surface markers are a class of PM proteins that are able to respond to or sense the environment around the cell and contain an extracellular domain, which has the benefit of being available for detection by an antibody.
What is cytoplasmic marker?
Cytoplasmic markers are key tools for enabling researchers to determine the location of novel proteins within the cell by reliability labeling the cytoplasm. The cytoplasm of cells consists of everything within the outer cell membrane. This includes the cytosol and organelles, but excludes the nucleus.
www.stressmarq.com/research/neuroscience/cell-structure/neuron-markers/cytoplasmic-markers/?v=7516fd43adaa
Hello Iman Aldybiat
You can try CD11b and CD66b as activated neutrophil markers.
Best.
Dear Iman Aldybiat ,
You may use above markers (CD11b, CD66b of CD63) on neutrophils for activation condition along with CD62L. (This CD62L will shade on activation).
All the best.
Malcolm Nobre Thank you for your reply,
from your experience ,how of these markers work for activated nucleus, more specifically (decondensed DNA) . Thanks
Jitendra Kumar Shandilya Thank you for your reply and for the options.
From your experience ,how of these markers work for activated nucleus, more specifically (decondensed DNA) . Thank you
Len Leonid Mizrah Thank you for your reply and for the useful infos regarding FACS
Activated neutrophil has higher CD11b expression than normal, but it's little complicated to using it as an activation marker.
(Article Neutrophil CD11B Expression and Neutrophil Activation in Pre-Eclampsia
)In mice, it has CD11b+ Ly6G+ (clone : only 1A8. RB6 also bind to Ly6C) character, and 'aged', not activated neutrophil also has lower CD62L level.
(Article Neutrophil ageing is regulated by the microbiome
)As I know, activation marker for neutrophil is not well elucidated yet, but I think I saw some unpublished data in seminar.
Geon Ho Bae what kind of complication you faced when using this marker? thank you
Iman Aldybiat I don't know whether it is depend on circadian cycle or something, some of blood neutrophils have CD11b^hi character in steady state mice.
So I think if experimental group has higher CD11b expression than control group, it could mean that neutrophils activated, but cannot rule out the possibility that another factor induced CD11b expression.
Geon Ho Bae Ok thanks for your answer. I am working on human samples. I will keep this notice in mind and will check whether there is difference or not.
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