Anti-acetyl-alpha tubulin Antibody, clone 6-11B-1

Microtubules are cytoskeletal filaments composed of of alpha-/beta-tubulin heterodimers that self-assemble head-to-tail to form protofilaments and laterally to form a hollow tube. Various post-translational modifications (PTMs), including acetylation, polyglutamylation, polyglycylation, detyrosination, phosphorylation, and palmitoylation, can regulate the polymerization properties of tubulins and/or their interactions with microtubule associated proteins (MAPs) and motor proteins. A subset of cytoplasmic microtubules and microtubules in the spindle, axon, and cilia are known to contain alpha-tubulin acetylated via the epsilon-amino group of Lysine-40 (K40). The GNAT lysine acetyltransferase family member MEC-17 (or alpha-tubulin acetyltransferase/alphaTAT) catalyzes K40 acetylation, while HDAC6 and sirtuin2 (SIRT2) are known to catalyze tubulin deacetylation. Experimental evidences indicate that K40 acetylation primes alpha-tubulin for further PTM(s), which cannot be reversed by later deactylation of K40ac. Subsequently, the acetylated and deacetylated alpha-tubulin in native microtubules are structurally distinct from that of unacetylated alpha-tubulin (never before has its K40 acetylated).

Experimental evidences indicate that the acetylated and deacetylated α-tubulin in native microtubules are structurally distinct from that of unacetylated α-tubulin, and that clone 6-11B-1 recognizes acetylated and deacetylated α-tubulin, but not unacetylated (never acetylated) α-tubulin. While 6-11B-1 immunostaining pattern resembles that of acetyl-Lys40 antibodies on normal cylcling cells, caution must be taken when interpreting immunostaining results using this mAb on cells subjected to tubulin deacetylation manipulations by exogenous deacetylase expression or via chemical means.

15 S dynein fraction from sea urchin sperm axoneme.

Research Category
Cell Structure

Research Sub Category
Adhesion (CAMs)

This Anti-acetyl-alpha tubulin Antibody, clone 6-11B-1 is validated for use in Western Blotting, Immunocytochemistry, Dot Blot and Electron Microscopy for the detection of acetyl-alpha tubulin.

Western Blotting Analysis: 4.0 µg/mL from a representative lot detected acetyl-alpha tubulin in 10 µg of mouse brain tissue lysate.
Immunocytochemistry Analysis: A representative lot detected acetylated alpha-tubulin in cultured normal human kidney epithelial cells (Courtesy of Dr Christopher Ward, University of Kansas Medical Center).
Western Blotting Analysis: A representative lot detected alpha-tubulin in sea urchin sperm axoneme lysates, but not in lysates from taxol-stabilized microtubules from sea urchin eggs (Piperno, G., and Fuller, M. (1985). J Cell Biol.101(6):2085-2094).
Dot Blot Analysis: A representative lot detected alpha-tubulin in both the ionic detergent Sarkosyl-soluble and insoluble fractions from sea urchin spern axoneme extracts (Piperno, G., and Fuller, M. (1985). J Cell Biol.101(6):2085-2094).
Immunocytochemistry Analysis: A representative lot detected acetylated and deacetylated, but not unacetylated alpha-tubulin in COS-7 green monkey kidney fibroblasts and Ptk2 rat kangaroo kidney epithelial cells by fluorescent immunocytochemistry (Soppina V., et al. (2012). PLoS One. 7(10):e48204).
Electron Microscopy Analysis: The Fab fragment of clone 6-11B-1 has been shown to stain Taxol-stabilized microtubules polymerized in vitro from purified bovine brain tubulin with or without SIRT2-catalyzed deacetylation (Soppina V., et al. (2012). PLoS One. 7(10):e48204).

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: 4.0 µg/mL of this antibody detected acetyl-alpha tubulin in 10 µg of HeLa cell lysate.

~50 kDa observed

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG2bκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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