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Figure 1: Immunoblot analysis of beta-tubulin protein glutamylation using anti-beta-Tubulin, mAb (AXO45) (Prod. No. AG-20B-0085). Method: HEK-293T cells are grown in standard culture conditions, transfected with plasmids expressing the different glutamylases, TTLL4 and TTLL6 and are run on a 10% SDS-PAGE (specific protocol to separate alpha- and beta-tubulin (Magiera & Janke, 2013)). The proteins are transferred to a nitrocellulose membrane and detected by standard immunoblot protocol using the AXO45 (1:1000) in PBS containing 0.1% Tween-20 for washing steps and 2.5% fat free milk for antibody incubation. In all the cell extracts, AXO45 specifically detects beta-tubulin, with no cross-reactivity to alpha-tubulin. Glutamylation added by either TTLL4 or TTLL6 appears to not affect this detection. Picture courtesy of Sudarshan Gadadhar, Shreyangi Chakraborty, Mariya Genova and Carsten Janke, Institut Curie, Orsay.
Figure 1: Immunoblot analysis of beta-tubulin protein glutamylation using anti-beta-Tubulin, mAb (AXO45) (Prod. No. AG-20B-0085). Method: HEK-293T cells are grown in standard culture conditions, transfected with plasmids expressing the different glutamylases, TTLL4 and TTLL6 and are run on a 10% SDS-PAGE (specific protocol to separate alpha- and beta-tubulin (Magiera & Janke, 2013)). The proteins are transferred to a nitrocellulose membrane and detected by standard immunoblot protocol using the AXO45 (1:1000) in PBS containing 0.1% Tween-20 for washing steps and 2.5% fat free milk for antibody incubation. In all the cell extracts, AXO45 specifically detects beta-tubulin, with no cross-reactivity to alpha-tubulin. Glutamylation added by either TTLL4 or TTLL6 appears to not affect this detection. Picture courtesy of Sudarshan Gadadhar, Shreyangi Chakraborty, Mariya Genova and Carsten Janke, Institut Curie, Orsay.
Figure 1: Immunoblot analysis of beta-tubulin protein glutamylation using anti-beta-Tubulin, mAb (AXO45) (Prod. No. AG-20B-0085). Method: HEK-293T cells are grown in standard culture conditions, transfected with plasmids expressing the different glutamylases, TTLL4 and TTLL6 and are run on a 10% SDS-PAGE (specific protocol to separate alpha- and beta-tubulin (Magiera & Janke, 2013)). The proteins are transferred to a nitrocellulose membrane and detected by standard immunoblot protocol using the AXO45 (1:1000) in PBS containing 0.1% Tween-20 for washing steps and 2.5% fat free milk for antibody incubation. In all the cell extracts, AXO45 specifically detects beta-tubulin, with no cross-reactivity to alpha-tubulin. Glutamylation added by either TTLL4 or TTLL6 appears to not affect this detection. Picture courtesy of Sudarshan Gadadhar, Shreyangi Chakraborty, Mariya Genova and Carsten Janke, Institut Curie, Orsay.

anti-beta-Tubulin, mAb (AXO45)

AG-20B-0085
AdipoGen Life Sciences
ApplicationsImmunoPrecipitation, Western Blot, ImmunoCytoChemistry, ImmunoHistoChemistry
Product group Antibodies
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Overview

  • Supplier
    AdipoGen Life Sciences
  • Product Name
    anti-beta-Tubulin, mAb (AXO45)
  • Delivery Days Customer
    10
  • Applications
    ImmunoPrecipitation, Western Blot, ImmunoCytoChemistry, ImmunoHistoChemistry
  • Certification
    Research Use Only
  • Clonality
    Monoclonal
  • Clone ID
    AXO45
  • Concentration
    1 mg/ml
  • Estimated Purity
    >95%
  • Host
    Mouse
  • Isotype
    IgG1
  • Scientific Description
    Monoclonal Antibody. Detects almost all isotypes of beta-tubulin specifically in cells and tissues of mouse, human and ciliates. Applications: ICC, IHC, IP and WB. Clone: AXO45. Isotype: Mouse IgG1. The beta-tubulin monoclonal antibody (AXO45) was developed by isolating the cilia from unicellular multiciliated Paramecium and purifying the core microtubule-based structures of the cilia, the axonemes. These axonemes were electrophoresed on a SDS-PAGE and the tubulin band was used as an antigen to develop monoclonal antibodies targeting both alpha- and beta-tubulin (A.M. Callen, et al.; 1994). The antibody AXO45 recognizes the C-terminal region between residues 257 and 442 of beta-tubulin. Although the antibody was raised against the beta-tubulin found in cilia, it is able to recognize beta-tubulin of all microtubule networks, indicating that it can potentially recognize different isotypes of beta-tubulin found in mammals. - The beta-tubulin monoclonal antibody (AXO45) was developed by isolating the cilia from unicellular multiciliated Paramecium and purifying the core microtubule-based structures of the cilia, the axonemes. These axonemes were electrophoresed on a SDS-PAGE and the tubulin band was used as an antigen to develop monoclonal antibodies targeting both alpha- and beta-tubulin (A.M. Callen, et al.; 1994). The antibody AXO45 recognizes the C-terminal region between residues 257 and 442 of beta-tubulin. Although the antibody was raised against the beta-tubulin found in cilia, it is able to recognize beta-tubulin of all microtubule networks, indicating that it can potentially recognize different isotypes of beta-tubulin found in mammals.
  • Storage Instruction
    -20°C,2°C to 8°C
  • UNSPSC
    12352203

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Figure 1: Immunoblot analysis of protein glutamylation with anti-beta-Tubulin (beta-monoE), pAb (IN115) (Prod. No. AG-25B-0039). Method: HEK-293T cells are grown in standard culture conditions, transfected with plasmids expressing the different glutamylases and are run on a 10% SDS-PAGE (specific protocol to separate alpha- and beta-tubulin (Magiera & Janke, 2013)). Tubulin purified from pork brain was used as a positive control for glutamylated tubulin. The proteins are transferred to a nitrocellulose membrane and detected by standard immunoblot protocol using the beta-monoE (1:5,000) in PBS containing 0.1% Tween-20 for washing steps and 2.5% fat free milk for antibody incubation. In untransfected HEK-293T cell extracts, no tubulin is detected, even on high exposure. After expression of TTLL4, predominantly beta-tubulin is detected with beta-monoE, but the antibody also faintly detects the heterogeneous mixture of different, yet unidentified substrates modified by TTLL4. In cells expressing TTLL5 and TTLL6, the beta-monoE detects only beta-tubulin, while expression of TTLL7 generates beta-monoE-positive alpha- and beta-tubulin. In the purified brain tubulin, the antibody specifically detects glutamylation of beta-tubulin. Picture courtesy of Sudarshan Gadadhar, Shreyangi Chakraborty, Mariya Genova and Carsten Janke, Institut Curie, Orsay.
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