Ab11309 specifically recognizes an epitope in the carboxy-terminal part of beta-tubulin (between amino acids 281-446).
Cultured chicken fibroblasts or BHK cells.
If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. This product or portions thereof is manufactured under license from Carnegie Mellon University under U.S. Patent Number 5,268,486 and related patents. Cy and CyDye are trademarks of GE Healthcare Limited. This material is also subject to proprietary rights of GE Healthcare Bio-Sciences Corp. and Carnegie Mellon University and made and sold under License from GE Healthcare Bio-Sciences Corp. This product is licensed for sale only for research. It is NOT licensed for any other use. There is no implied license hereunder for any commercial use. COMMERCIAL USE shall include: 1 Sale, lease, license or other transfer of the material or any material derived or produced from it. 2 Sale, lease, license or other grant of rights to use this material or any material derived or produced from it. 3 Use of this material to perform services for a fee for third parties. If you require a commercial license to use this material and do not have one, please return this material, unopened to Abcam Plc of 330 Cambridge Science Park, Cambridge, CB4 0FL, and any money paid for the material will be refunded.
The product is prepared by conjugation of Cy3 to purified monoclonal anti-beta-tubulin antibody. The conjugate is purified by gel filtration to remove unbound Cy3 fluorophore. F/P Molar Ratio: (Cy3:Ab) 3 to 9.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent dilution. Predicted molecular weight: 50 kDa.
1/100. This dilution was determined by direct immunofluorescent labeling of cultured chicken fibroblasts or BHK cells. The conjugate may be used in double labeling experiments with fluorescein tagged antibodies.
Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain.
Ubiquitously expressed with highest levels in spleen, thymus and immature brain.
Cortical dysplasia, complex, with other brain malformations 6 Skin creases, congenital symmetric circumferential, 1
Belongs to the tubulin family.
The highly acidic C-terminal region may bind cations such as calcium.
Some glutamate residues at the C-terminus are polyglutamylated, resulting in polyglutamate chains on the gamma-carboxyl group (PubMed:26875866). Polyglutamylation plays a key role in microtubule severing by spastin (SPAST). SPAST preferentially recognizes and acts on microtubules decorated with short polyglutamate tails: severing activity by SPAST increases as the number of glutamates per tubulin rises from one to eight, but decreases beyond this glutamylation threshold (PubMed:26875866). Some glutamate residues at the C-terminus are monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella). Both polyglutamylation and monoglycylation can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of monoglycylation is still unclear. Phosphorylated on Ser-172 by CDK1 during the cell cycle, from metaphase to telophase, but not in interphase. This phosphorylation inhibits tubulin incorporation into microtubules.
Immunocytochemistry/ Immunofluorescence - beta Tubulin antibody [TUB 2.1] (Cy3 ®) (ab11309)This image is courtesy of an anonymous Abreview.
ab11309 staining beta Tubulin in Mouse cultured cortical neurons by Immunocytochemistry/Immunofluorescence. Cells were plated on a poly-L-Lysine coated cover slip before being washed twice with PBS and fixed with 4% PFA at room temperature. Cells were permeabilized in 0.5%(w/v) saponin in PBS prior to blocking in 10% fetal calf serum in PBS-saponin for 2 hours at 23°C. The primary antibody was diluted 1/1000 in the blocking buffer and incubated with the sample for 2 hours at 23°C. After additional washing cover splips were mounted on glass slides with ProLong® Gold antifade reagent with DAPI (Invitrogen). The image was acquired with a 40x objective.
Lysis Buffer was 1% Triton X-100, 20mM TrisHCl pH 7.5, 10mM EGTA, 150mM NaCl, protease inhibitors. The image was acquired by scanning the blot with a FLA-9000 Starion laserscanner at 300V and 200µm resolution. The above mentioned exposure time is just the time it took to scan the blot but does not correlate to signal intensity.