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anti-β-Tubulin (β-monoE), pAb (IN115)

Description
Polyglutamylation is an evolutionarily conserved protein post-translational modification in which glutamate side chains of variable lengths are formed on the modified protein. The most prominent substrate is tubulin, the building block of microtubules (MTs). Polyglutamylation has been found on both, α- and β-tubulin. It is catalyzed by multiple enzymes that belong to the family of tubulin tyrosine-ligase like (TTLL) enzymes. These TTLLs have substrate (α- vs β-tubulin) as well as reaction (initiation vs elongation) specificities. These subtle changes in levels of polyglutamylation on specific tubulin isoforms could in turn be influencing the diverse MT functions that are regulated by polyglutamylation. The β-monoE antibody selectively labels glutamylation of β-tubulin due to its specificity to a sequence motif (GE(*-E)F) that only exists in β-tubulin isotypes. It allows to specifically detect β-tubulin glutamylation in brain tubulin, where other antibodies predominantly detect α-tubulin glutamylation.

Product References
Distinct roles of alpha- and beta-tubulin polyglutamylation in controlling axonal transport and in neurodegeneration: S. Bodakuntla, et al.; EMBO J. 40, e108498 (2021)

List Print
Product Details
Product Type Polyclonal Antibody
Properties
Source/Host Rabbit
Immunogen/Antigen Synthetic peptide corresponding to D431EQGEFE(E-COOH*)EEEG441-NH2 of human tubulin β-2A chain (*secondary glutamate branched from γ-carboxyl group of glutamate as isopeptide bond).
Application

Western Blot: (1:2000)

Immunocytochemistry: (1:5000)

Immunohistochemistry

Immunoprecipitation

Optimal conditions must be determined individually for each application.
Crossreactivity Human
Mouse
Specificity

Recognizes the posttranslational modification glutamylation with specific reactivity for the GE(*-E)F motive. Because the sequence GEF is specific to several β-tubulin isotypes, the antibody selectively detects mono-glutamylated β-tubulin.
Purity ≥95% (SDS-PAGE)
Purity Detail Epitope-affinity purified.
Concentration 1mg/ml
Formulation Liquid. In PBS containing 0.02% Proclin 300.
Isotype Negative Control

Rabbit IgG

Other Product Data

Recognizes predominantly mono-glutamylated β-tubulin. However, in cells overexpressing enzymes that can modify both α- and β-tubulin, it might detect glutamylation on α-tubulin as well. Requires GE(*-E)F motive, which is present in several β-tubulin isotypes. No other specificity to particular tubulin isotypes nor to tubulin from particular species are observed. The use of the antibody at too high concentrations obscures its specificity in immunofluorescence. 
Accession Number Q13885
Shipping and Handling
Shipping BLUE ICE
Short Term Storage +4°C
Long Term Storage -20°C
Handling Advice After opening, prepare aliquots and store at -20°C.
Avoid freeze/thaw cycles.
Use/Stability Stable for at least 1 year after receipt when stored at -20°C.

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