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Glu-Tubulin antibody - 302 011

alpha-Tubulin is a major cytoskeleton protein
Mouse monoclonal purified IgG
Cat. No.: 302 011
Amount: 100 µg
Price: $415.00
Cat. No. 302 011 100 µg purified IgG, lyophilized. Albumin and azide were added for stabilization. For reconstitution add 100 µl H2O to get a 1mg/ml solution in PBS. Then aliquot and store at -20°C to -80°C until use.
Antibodies should be stored at +4°C when still lyophilized. Do not freeze!
Applications
 
WB: 1 : 1000 (AP staining) (see remarks) gallery  
IP: not tested yet
ICC: 1 : 200 up to 1 : 500 gallery  
IHC: 1 : 500 gallery  
IHC-P: 1 : 1000 gallery  
ELISA: yes

Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section.

Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section.

Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section.

Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section.

Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.

Enzyme-linked immunosorbent assay (ELISA); a frequently employed method to quantify target-molecules in solution. The detection of some proteins requires special solubilization steps. For further information, please refer to the „Remarks“ section.

Clone 1D5
Subtype IgG1 (κ light chain)
Immunogen Synthetic peptide corresponding to residues near the c-terminus of human Glu-α-tubulin. (UniProt Id: Q71U36)
Epitop AA 448 to 450 from human Glu-α-tubulin (UniProt Id: Q71U36)
Reactivity Reacts with: human (Q71U36), rat (P68370), mouse (P68369), zebrafish, eukaryotes, other vertebrates.
Other species not tested yet.

Detects also cilia of Paramecium.
Specificity Specific for detyrosinated α-tubulin (glu-tubulin) and polyglutamylated tubulin (also β-tubulin). No cross reaction to tyrosinated tubulin.
Remarks

WB: Tween 20 (other detergents not yet tested) should not be added to the blocking and washing solution. It greatly diminishes the intensity of the signal.

Data sheet 302_011.pdf

References for Glu-Tubulin - 302 011

Tubulin detyrosination promotes monolayer formation and apical trafficking in epithelial cells.
Zink S, Grosse L, Freikamp A, Bänfer S, Müksch F, Jacob R
Journal of cell science (2012) 125Pt 24: 5998-6008. 302 011 WB, ICC
A family of protein-deglutamylating enzymes associated with neurodegeneration.
Rogowski K, van Dijk J, Magiera MM, Bosc C, Deloulme JC, Bosson A, Peris L, Gold ND, Lacroix B, Bosch Grau M, Bec N, et al.
Cell (2010) 1434: 564-78. 302 011 WB
Turnover of the carboxy-terminal tyrosine of alpha-tubulin and means of reaching elevated levels of detyrosination in living cells.
Wehland J, Weber K
Journal of cell science (1987) 88 ( Pt 2): 185-203. 302 011 WB, ICC
SF-Assemblin genes in Paramecium: phylogeny and phenotypes of RNAi silencing on the ciliary-striated rootlets and surface organization.
Nabi A, Yano J, Valentine MS, Picariello T, Van Houten JL
Cilia (2019) 8: 2. 302 011 ICC
The Ciliary Protein IFT57 in the Macronucleus of Paramecium.
Shi L, Koll F, Arnaiz O, Cohen J
The Journal of eukaryotic microbiology (2017) : . 302 011 ICC
BDNF/trkB Induction of Calcium Transients through Cav2.2 Calcium Channels in Motoneurons Corresponds to F-actin Assembly and Growth Cone Formation on β2-Chain Laminin (221).
Dombert B, Balk S, Lüningschrör P, Moradi M, Sivadasan R, Saal-Bauernschubert L, Jablonka S
Frontiers in molecular neuroscience (2017) 10: 346. 302 011 ICC; tested species: mouse
Kif26b controls endothelial cell polarity through the Dishevelled/Daam1-dependent planar cell polarity-signaling pathway.
Guillabert-Gourgues A, Jaspard-Vinassa B, Bats ML, Sewduth RN, Franzl N, Peghaire C, Jeanningros S, Moreau C, Roux E, Larrieu-Lahargue F, Dufourcq P, et al.
Molecular biology of the cell (2016) 276: 941-53. 302 011 ICC
Mechanisms for axon maintenance and plasticity in motoneurons: alterations in motoneuron disease.
Jablonka S, Dombert B, Asan E, Sendtner M
Journal of anatomy (2014) 2241: 3-14. 302 011 ICC
Reduction of meckelin leads to general loss of cilia, ciliary microtubule misalignment and distorted cell surface organization.
Picariello T, Valentine MS, Yano J, Van Houten J
Cilia (2014) 31: 2. 302 011 ICC
Tubulin detyrosination promotes monolayer formation and apical trafficking in epithelial cells.
Zink S, Grosse L, Freikamp A, Bänfer S, Müksch F, Jacob R
Journal of cell science (2012) 125Pt 24: 5998-6008. 302 011 WB, ICC
Hypoxia stimulates carcinoma invasion by stabilizing microtubules and promoting the Rab11 trafficking of the alpha6beta4 integrin.
Yoon SO, Shin S, Mercurio AM
Cancer research (2005) 657: 2761-9. 302 011 ICC
Turnover of the carboxy-terminal tyrosine of alpha-tubulin and means of reaching elevated levels of detyrosination in living cells.
Wehland J, Weber K
Journal of cell science (1987) 88 ( Pt 2): 185-203. 302 011 WB, ICC
Monocilia on chicken embryonic endocardium in low shear stress areas.
Van der Heiden K, Groenendijk BC, Hierck BP, Hogers B, Koerten HK, Mommaas AM, Gittenberger-de Groot AC, Poelmann RE
Developmental dynamics : an official publication of the American Association of Anatomists (2006) 2351: 19-28. 302 011 IHC
Tubulin tyrosine ligase structure reveals adaptation of an ancient fold to bind and modify tubulin.
Szyk A, Deaconescu AM, Piszczek G, Roll-Mecak A
Nature structural & molecular biology (2011) 1811: 1250-8. 302 011
Structural basis of microtubule severing by the hereditary spastic paraplegia protein spastin.
Roll-Mecak A, Vale RD
Nature (2008) 4517176: 363-7. 302 011
Two catalytic domains are required for protein deacetylation.
Zhang Y, Gilquin B, Khochbin S, Matthias P
The Journal of biological chemistry (2006) 2815: 2401-4. 302 011
Coordination of microtubule and microfilament dynamics by Drosophila Rho1, Spire and Cappuccino.
Rosales-Nieves AE, Johndrow JE, Keller LC, Magie CR, Pinto-Santini DM, Parkhurst SM
Nature cell biology (2006) 84: 367-76. 302 011
Monoclonal antibody ID5: epitope characterization and minimal requirements for the recognition of polyglutamylated alpha- and beta-tubulin.
Rüdiger AH, Rüdiger M, Wehland J, Weber K
European journal of cell biology (1999) 781: 15-20. 302 011
Primary cilia in cultured mammalian cells: detection with an antibody against detyrosinated alpha-tubulin (ID5) and by electron microscopy.
Wheatley DN, Feilen EM, Yin Z, Wheatley SP
Journal of submicroscopic cytology and pathology (1994) 261: 91-102. 302 011
Cat. No.: 302 011
Amount: 100 µg
Price: $415.00
Tubulin detyrosination promotes monolayer formation and apical trafficking in epithelial cells.
Zink S, Grosse L, Freikamp A, Bänfer S, Müksch F, Jacob R
Journal of cell science (2012) 125Pt 24: 5998-6008. 302 011 WB, ICC
A family of protein-deglutamylating enzymes associated with neurodegeneration.
Rogowski K, van Dijk J, Magiera MM, Bosc C, Deloulme JC, Bosson A, Peris L, Gold ND, Lacroix B, Bosch Grau M, Bec N, et al.
Cell (2010) 1434: 564-78. 302 011 WB
Turnover of the carboxy-terminal tyrosine of alpha-tubulin and means of reaching elevated levels of detyrosination in living cells.
Wehland J, Weber K
Journal of cell science (1987) 88 ( Pt 2): 185-203. 302 011 WB, ICC
SF-Assemblin genes in Paramecium: phylogeny and phenotypes of RNAi silencing on the ciliary-striated rootlets and surface organization.
Nabi A, Yano J, Valentine MS, Picariello T, Van Houten JL
Cilia (2019) 8: 2. 302 011 ICC
The Ciliary Protein IFT57 in the Macronucleus of Paramecium.
Shi L, Koll F, Arnaiz O, Cohen J
The Journal of eukaryotic microbiology (2017) : . 302 011 ICC
BDNF/trkB Induction of Calcium Transients through Cav2.2 Calcium Channels in Motoneurons Corresponds to F-actin Assembly and Growth Cone Formation on β2-Chain Laminin (221).
Dombert B, Balk S, Lüningschrör P, Moradi M, Sivadasan R, Saal-Bauernschubert L, Jablonka S
Frontiers in molecular neuroscience (2017) 10: 346. 302 011 ICC; tested species: mouse
Kif26b controls endothelial cell polarity through the Dishevelled/Daam1-dependent planar cell polarity-signaling pathway.
Guillabert-Gourgues A, Jaspard-Vinassa B, Bats ML, Sewduth RN, Franzl N, Peghaire C, Jeanningros S, Moreau C, Roux E, Larrieu-Lahargue F, Dufourcq P, et al.
Molecular biology of the cell (2016) 276: 941-53. 302 011 ICC
Mechanisms for axon maintenance and plasticity in motoneurons: alterations in motoneuron disease.
Jablonka S, Dombert B, Asan E, Sendtner M
Journal of anatomy (2014) 2241: 3-14. 302 011 ICC
Reduction of meckelin leads to general loss of cilia, ciliary microtubule misalignment and distorted cell surface organization.
Picariello T, Valentine MS, Yano J, Van Houten J
Cilia (2014) 31: 2. 302 011 ICC
Tubulin detyrosination promotes monolayer formation and apical trafficking in epithelial cells.
Zink S, Grosse L, Freikamp A, Bänfer S, Müksch F, Jacob R
Journal of cell science (2012) 125Pt 24: 5998-6008. 302 011 WB, ICC
Hypoxia stimulates carcinoma invasion by stabilizing microtubules and promoting the Rab11 trafficking of the alpha6beta4 integrin.
Yoon SO, Shin S, Mercurio AM
Cancer research (2005) 657: 2761-9. 302 011 ICC
Turnover of the carboxy-terminal tyrosine of alpha-tubulin and means of reaching elevated levels of detyrosination in living cells.
Wehland J, Weber K
Journal of cell science (1987) 88 ( Pt 2): 185-203. 302 011 WB, ICC
Monocilia on chicken embryonic endocardium in low shear stress areas.
Van der Heiden K, Groenendijk BC, Hierck BP, Hogers B, Koerten HK, Mommaas AM, Gittenberger-de Groot AC, Poelmann RE
Developmental dynamics : an official publication of the American Association of Anatomists (2006) 2351: 19-28. 302 011 IHC
Tubulin tyrosine ligase structure reveals adaptation of an ancient fold to bind and modify tubulin.
Szyk A, Deaconescu AM, Piszczek G, Roll-Mecak A
Nature structural & molecular biology (2011) 1811: 1250-8. 302 011
Structural basis of microtubule severing by the hereditary spastic paraplegia protein spastin.
Roll-Mecak A, Vale RD
Nature (2008) 4517176: 363-7. 302 011
Two catalytic domains are required for protein deacetylation.
Zhang Y, Gilquin B, Khochbin S, Matthias P
The Journal of biological chemistry (2006) 2815: 2401-4. 302 011
Coordination of microtubule and microfilament dynamics by Drosophila Rho1, Spire and Cappuccino.
Rosales-Nieves AE, Johndrow JE, Keller LC, Magie CR, Pinto-Santini DM, Parkhurst SM
Nature cell biology (2006) 84: 367-76. 302 011
Monoclonal antibody ID5: epitope characterization and minimal requirements for the recognition of polyglutamylated alpha- and beta-tubulin.
Rüdiger AH, Rüdiger M, Wehland J, Weber K
European journal of cell biology (1999) 781: 15-20. 302 011
Primary cilia in cultured mammalian cells: detection with an antibody against detyrosinated alpha-tubulin (ID5) and by electron microscopy.
Wheatley DN, Feilen EM, Yin Z, Wheatley SP
Journal of submicroscopic cytology and pathology (1994) 261: 91-102. 302 011
Background

Microtubules are involved in a wide variety of cellular activities ranging from mitosis and transport events to cell movement and the maintainance of cell shape.
Tubulin itself is a globular protein which consists of two polypeptides, α-tubulin and β-tubulin. α- and β-tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter. Tyrosine ligase ads a C-terminal tyrosin to monomeric α-tubulin.
Assembled microtubules can again be detyrosinated by a cytoskeleton associated carboxypeptidase. Detyrosinated α-tubulin is referred to as Glu-α-tubulin. Another post-translational modification of detyrosinated α-tubulin is C-terminal polyglutamylation which is characteristic for microtubules in neuronal cells and the mitotic spindle. A third variant of detyrosinated α-tubulin is Δ2-tubulin which lacks the C-terminal glutamic acid. It cannot be tyrosinated by tyrosine ligase and is one of the dominant α-tubulin isoforms in neurons.