Confocal immunofluorescent analysis of HeLa cells, serum-starved (left) or 20% serum-treated (right), using β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) (red) and Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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Cell Signaling Technology antibody is conjugated to Alexa Fluor® 555 fluorescent dye and tested in-house for direct immunofluorescent analysis of human and monkey cells. The unconjugated antibody #2128 reacts with human, mouse, rat, monkey, bovine, zebrafish and fly β-tubulin protein. CST expects that β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) will also recognize β-tubulin in these species.
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
β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) detects endogenous levels of total β-tubulin protein.
Human, Mouse, Rat, Monkey, Zebrafish, Bovine
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to the amino terminus of human β-tubulin. The antibody was conjugated to Alexa Fluor® 555 under optimal conditions with an F/P ratio of 2-6.
The cytoskeleton consists of three types of cytosolic fibers: microtubules, microfilaments (actin filaments), and intermediate filaments. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. γ-tubulin is required to nucleate polymerization of tubulin subunits to form microtubule polymers. Many cell movements are mediated by microtubule action, including the beating of cilia and flagella, cytoplasmic transport of membrane vesicles, chromosome alignment during meiosis/mitosis, and nerve-cell axon migration. These movements result from competitive microtubule polymerization and depolymerization or through the actions of microtubule motor proteins (1).
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