In general, yes, they typically work. However, there are some limitations. Flow cytometry relies on the density of the antigen, and the cells are individually "scanned" for the fluorescent antibodies--individual excitation and emission in a light-sealed environment. Microscopes don't work that way--excitation and emission are not usually that focused--so your signal may be lower. If your antigen is not highly expressed on your cells (low/dim in flow cytometry), you will also likely have trouble seeing it with microscopy. One thing you can do to amplify your signal is use a biotinylated primary antibody, followed by a streptavidin-fluorophore conjugate. I think I once read that approximately 4 streptavidin molecules (they tend to form a tetramer) can bind to a biotinylated antibody--and the exact number isn't critical; the point is that it dramatically increases your signal. You can also use an unconjugated primary antibody followed by a biotinylated secondary antibody and THEN a streptavidin-fluorophore.
Many companies that sell antibodies will list what it has been validated for in terms of techniques. If your company does not, find out what clone (if mAb) and see if another company does list that antibody for your application. It's worth a phone call to their tech support.
Possibly, but the only way to know is to try it. The only thechnical limit I would foresee is having the right lamp and filters for the scope. Also, you may have to tweak your staining protocol, particularly fixation and premeabilization (if intracellular).
In general, yes, they typically work. However, there are some limitations. Flow cytometry relies on the density of the antigen, and the cells are individually "scanned" for the fluorescent antibodies--individual excitation and emission in a light-sealed environment. Microscopes don't work that way--excitation and emission are not usually that focused--so your signal may be lower. If your antigen is not highly expressed on your cells (low/dim in flow cytometry), you will also likely have trouble seeing it with microscopy. One thing you can do to amplify your signal is use a biotinylated primary antibody, followed by a streptavidin-fluorophore conjugate. I think I once read that approximately 4 streptavidin molecules (they tend to form a tetramer) can bind to a biotinylated antibody--and the exact number isn't critical; the point is that it dramatically increases your signal. You can also use an unconjugated primary antibody followed by a biotinylated secondary antibody and THEN a streptavidin-fluorophore.
Many companies that sell antibodies will list what it has been validated for in terms of techniques. If your company does not, find out what clone (if mAb) and see if another company does list that antibody for your application. It's worth a phone call to their tech support.
The CD130 molecule is expressed at low levels on almost all cell types , including a subset of CD4+ T lymphocytes , most monocytes , basophils , endothelial cells, epithelial cells , fibroblasts , hepatocytes and neural cells . It is expressed at high levels on activated and EBV-transformed B cells . It is not expressed on resting B cells , NK cells, nor is it expressed on neutrophils or dendritic cells .
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