Product Pathways - Cytoskeletal Signaling
Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb #5335
|5335S||100 µl (10 western blots)||---||In Stock||---|
|5335P||40 µl (4 western blots)||---||In Stock||---|
|5335||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Rat, Monkey, Zebrafish||Endogenous||52||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Species predicted to react based on 100% sequence homology: Xenopus.
Specificity / Sensitivity
Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb detects endogenous levels of α-tubulin only when acetylated at Lys40. This amino acid is not conserved in β-tubulin.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic acetylpeptide corresponding to residues surrounding Lys40 of human α-tubulin.
Western blot analysis of extracts from HeLa cells, untreated or TSA-treated (400 nM for 16 hours), using Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb (upper) and α-Tubulin (11H10) Rabbit mAb #2125 (lower). The acetyl-specificity of the antibody was verified by blocking with an acetyl- or non-acetylpeptide.
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A #9950 (right), using Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A (TSA) #9950 (1uM, 16 hours) using Acetyl-alpha-Tubulin (Lys40) (D20G3) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.
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).
The Elongator complex catalytic subunit (Elp3) acetylates α-tubulin at Lys40 while the histone deacetylase HDAC6 functions as a tubulin deacetylase. This post-transcriptional modification may be required for dynamic cell shape remodeling, cell motility, tubulin stability and terminal branching of cortical neurons (2,3).
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.