I want to do Ex Vivo Functional Analysis of human Dendritic Cells.
What i found was here:
1,Measurement of Export to the Cytosol in Dendritic Cells Using a Cytofluorimetry-Based Assay (Principal of the assay. Cells are loaded with CCF4 dye. Cells are then incubated with β-lactamase during different time points. If the β-lactamase undergoes export to the cytosol, the CCF4 dye is cleaved resulting in loss of FRET and the cells switch from 535 nm emission fluorescence (A) to 450 nm fluorescence (B) )
2,Analysis of Intracellular Trafficking of Dendritic Cell Receptors for Antigen Targeting (like the first one)
3,Cross-Presentation Assay for Human Dendritic Cells (Assessment of T Cell Clone Activation )
4,Characterization of Dendritic Cell Subsets Through Gene Expression Analysis
any other choice?
Hi,
You could also stimulate them with TLR agonist (LPS is the strongest) and look for activation markers up-regulation (CD86, CD80, CD83, CD40, HLA DR ...) and cytokine production (Type-I IFN, IL12p70, IP10 ... ).
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