Product Pathways - Cytoskeletal Signaling
Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb #5335
PhosphoSitePlus® protein, site, and accession data: TUBA1B
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC | H M R Mk Z | Endogenous | 52 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Z=Zebrafish
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
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 Blotting
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.
IF-IC
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).
Background
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).
- Westermann, S. and Weber, K. (2003) Nat. Rev. Mol. Cell Biol. 4, 938 -947.
- Creppe, C. et al. (2009) Cell 136, 551-564.
- Hubbert, C. et al. (2002) Nature 417, 455-458.
Application References
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Companion Products
- 3971 Acetyl-α-Tubulin (Lys40) Antibody
- 2125 α-Tubulin (11H10) Rabbit mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7075 Anti-biotin, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
- 9998 BSA
- 9999 Nonfat Dry Milk
For Research Use Only. Not For Use In Diagnostic Procedures.
