We are trying to conduct an experiment utilizing CyTOF. We are wondering if there is a protocol for fixation and permeabilization that will allow us to stain for both phosphorylated and non-phosphorylated proteins. Typically we would use methanol for the phosphorylated proteins and the Foxp3 perm-fix buffer for non-phosphorylated proteins.
Jitendra Kumar Shandilya
For flowcytometry, technically it is possible with choice of antibodies. One need to choose antibodies against two different domains (one for phosphorylation domain and other for a domain not near to the first). Methanol would work for this protocol. You will be able to look at total protein as well as phosphorylation out of total.
Not much familiar about mass cytometry experiment, so no idea about it.
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