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Problem: Lack of Staining
Possible Source
Lack of antigen
Test or Action
Check protein expression by in situ hybridization (in some rare cases translation may be blocked even though mRNA is detected)
Possible Source
Antibodies do not work due to improper storage
Test or Action
Aliquot antibodies into smaller volumes sufficient to make a working solution for a single experiment. Store aliquots in a freezer (-20 to -70°C) and avoid repeated freeze-thaw cycles