Anti-alpha-Tubulin Antibody, clone YOL1/34

General description

Tubulin alpha-1 chain (UniProt P09733; also known as alpha-tubulin) is encoded by the TUB1 gene (Gene ID 854889) in yeast. Microtubules are dynamic protein filaments involved in a wide variety of cellular activities ranging from mitosis and transport events to cell movement and the maintenance of cell shape. Microtubules are assembled from α- and β-tubulin subunits, both of which are subjected to various post-translational modifications (PTMs), including acetylation, polyglutamylation, polyglycylation, detyrosination, phosphorylation, and palmitoylation. These PTMs can regulate the polymerization properties of tubulins and/or their interactions with microtubule associated proteins (MAPs) and motor proteins. Because the C-terminus tyrosine/phenylalanine is preceded by glutamate (E) residues, the detyrosinated/dephenylalaninated α-tubulin is also referred to as Glu-tubulin. While tyrosine can be reattached to the C-terminus of the Glu-tubulin by tubulin-tyrosine ligase in mammals, phenylalanine is not added posttranslationally to the Glu-tubulin in yeast despite the presence of the yeast YBR094w gene that exhibits significant homology to tubulin-tyrosine ligase in other organisms

Specificity

Clone YOL1/34 reacts with alpha-tubulin, but not beta-tubulin, by targeting a C-terminal epitope highly conserved among alpha-tubulin subtypes. Epitope is not present in alpha-tubulin 4B (TUBA4B; UniProt Q9H853).

Expected to react with a broad range of species based on 100% sequence homology.

Immunogen

Yeast tubulin (Kilmartin, J.V., et al. (1982). J. Cell Biol. 93(3):576-582).

Application

Immunocytochemistry Analysis: A 1:200 dilution from a representative lot detected alpha-tubulin in HeLa cell lysate.

Western Blotting Analysis: 1 µg/mL from a representative lot detected alpha-tubulin in 10 µg of NIH/3T3 mouse fibroblast and PC12 rat pheochromocytoma cell lysates.

Affinity Binding Assay: Pig brain α-tubulin, α-tubulin/β-tubulin heterodimer, as well as α-tubulin proteolytic fragments a.a. 414-425 and a.a. 403-422, but not β-tubulin or actin, effectively competed against immobilized α-tubulin for YOL 1/34 binding (Breitling, F., and Little, M. (1986). J. Biol. Chem. 189(2):367-370).

Electron Microscopy Analysis: A representative lot immunostained microtubules of crude yeast nuclear envelope preparation (Kilmartin, J.V., et al. (1982). J. Cell Biol. 93(3):576-582).

ELISA Analysis: A representative lot detected pig brain α-tubulin by direct (non-sandwich) ELISA (Breitling, F., and Little, M. (1986). J. Biol. Chem. 189(2):367-370).

Immunocytochemistry Analysis: A representative lot immunostained fixed NIH/3T3 cells as well as yeast spheroplasts and nuclei preparations (Kilmartin, J.V., et al. (1982). J. Cell Biol. 93(3):576-582).

Immunocytochemistry Analysis: A representative lot immunostained microtubules of fixed Dictyostelium (amoeba) as well as isolated and stabilized amoeba cytoskeleton (Kilmartin, J.V., et al. (1982). J. Cell Biol. 93(3):576-582).

Western Blotting Analysis: A representative lot detected α-tubulin in spermatozoa sample from from fertile and infertile men (Breitling, F., et al. (1991). Gene. 104(2):147-153).

This rat monoclonal Anti-alpha-Tubulin Antibody, clone YOL1/34, Cat. No. CBL270-I is validated for use in Affinity Binding Assay, Electron Microscopy, ELISA, Immunocytochemistry, and Western Blotting for the detection of alpha-tubulin.

Quality

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: 1 µg/mL of this antibody detected alpha-tubulin in 10 µg of HeLa cell lysate.

Target description

~50 kDa observed. 50.14/46.30 kDa (human alpha-1A isoform 1/2; TUBA1A; TUBA3), 50.15/37.22 kDa (human alpha-1B isoform 1/2; TUBA1B), 44.96 kDa (human alpha-2; TUBA2; TUBA3C; TUBA3D), 49.92/48.33 kDa (human alpha-4A isoform 1/2; TUBA4A; TUBA1), 50.14 kDa (mouse & rat alpha-1A; Tuba1a; Tuba1), 50.15 kDa (mouse & rat alpha-1B; Tuba1b; Tuba2), 49.96 kDa (mouse & rat alpha-3; Tuba3; Tuba3a; Tuba3b), 49.92 kDa (mouse & rat alpha-4A; Tuba4a; Tuba4), 49.80 kDa (yeast) calculated. Uncharacterized bands may be observed in some lysate(s).

Linkage

Replaces: CBL270

Physical form

Format: Purified

Purified rat IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Other Notes

Concentration: Please refer to lot specific datasheet.