Confocal immunofluorescent analysis of COS cells transiently transfected with a Myc-tagged fusion protein using Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 555 Conjugate) (red) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 488 Conjugate) #3623 (green).
This Cell Signaling Technology antibody was conjugated to Alexa Fluor® 555 fluorescent dye and tested in-house for immunofluorescence in cells transfected with Myc-tagged protein.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 555 Conjugate) detects transfected proteins containing the Myc epitope tag. The antibody recognizes the Myc-tag fused to either the amino or carboxy terminus of targeted proteins in transfected mammalian cells.
All Species Expected
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues 410-419 of human c-Myc (EQKLISEEDL). The antibody was conjugated to Alexa Fluor® 555 under optimal conditions with an F/P ratio of 2-6.
Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein’s biochemical properties.
The Myc epitope tag is widely used to detect expression of recombinant proteins in bacteria, yeast, insect and mammalian cell systems (1).