Technical Data
T9154-04
Tubulin
Description:
Tubulins, major components of the cellular cytoskeleton, form microtubules that are critical for cellular structure, migration, cell division and intracellular trafficking. There are at least three types of tubulins, alpha, beta and gamma. Each has a molecular weight of approximately 55kD. Microtubules are large structures composed of alternating alpha and beta subunits. Compounds that block microtubule formation, such as Taxol and the vinca alkaloids, inhibit cell division and are effective anti-cancer agents. The epitope recognized by this antibody has been extensively studied and would appear to be a linear sequence requiring an aromatic residue at the C-terminus, with the two adjacent amino acids being negatively charged (represented by Gly-Gly-Tyr in Tyr-Tubulin). The antibody has been used in epitope tagging procedures to detect proteins tagged with a C-terminal Gly-Gly-Phe epitope. These sequence requirements have been reported to result in some cross-reactivity with other proteins in certain circumstances, including E. coli rec A and oxidized actin.

Applications:
Suitable for use in RIA, ELISA, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
ELISA: 1:100-1:1000
Western Blot: 1:500. Detects tubulin on 10ug of A431 lysates. A431 lysate was resolved by electrophoresis, transferred to PVDF membrane and probed with T9154-04 (1:500). Proteins were visualizes using a goat anti-rat secondary antibody conjugated to HRP and a chemiluminescence detection system.
Immunohistochemistry (Frozen): 1:50-1:100
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.Tubulins, major components of the cellular cytoskeleton, form microtubules that are critical for cellular structure, migration, cell division and intracellular trafficking. There are at least three types of tubulins, alpha, beta and gamma. Each has a molecular weight of approximately 55kD. Microtubules are large structures (300 angstrom diameter) composed of alternating alpha and beta subunits. Compounds that block microtubule formation, such as Taxol and the vinca alkaloids, inhibit cell division and are effective anti-cancer agents. The epitope recognized by this antibody has been extensively studied and would appear to be a linear sequence requiring an aromatic residue at the C-terminus, with the two adjacent amino acids being negatively charged (represented by Gly-Gly-Tyr in Tyr-Tubulin). The antibody has been used in epitope tagging procedures to detect proteins tagged with a C-terminal Gly-Gly-Phe epitope. These sequence requirements have been reported to result in some cross-reactivity with other proteins in certain circumstances, including E. coli rec A and oxidized actin.

Applications:
Suitable for use in RIA, ELISA, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
ELISA: 1:100-1:1000
Western Blot: 1:500. Detects tubulin on 10ug of A431 lysates. A431 lysate was resolved by electrophoresis, transferred to PVDF membrane and probed with T9154-04 (1:500). Proteins were visualizes using a goat anti-rat secondary antibody conjugated to HRP and a chemiluminescence detection system.
Immunohistochemistry (Frozen): 1:50-1:100
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG2a2Q1105Affinity Purified
SizeStorageShippingSourceHost
100ug4C (-20C Glycerol)Blue IceRat
Concentration:
~1mg/ml
Immunogen:
Yeast tubulin
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide.
Specificity:
Recognizes the alpha subunit of human tubulin, specificity binding to so called Tyr-Tubulin at ~55kD, which is produced by post translational modification of tubulin. Species crossreactivity: mouse and rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Kilmartin, J. V. et al. (1982). Rat monoclonal anti-tubulin antibodies derived by using a new non-secreting rat cell line. J. Cell. Biol. 93:576582. 2. Skinner, R.H. et al. (1991). Use of the Glu-Glu-Phe C-terminal epitope for rapid purification of the catalytic domain of normal and mutant ras GTPase-activating proteins. J. Biol. Chem. 266:1416314166. 3. Wehland, J. et al. (1984). Amino acid sequence requirements in the epitope recognized by the alpha tubulin specific rat monoclonal antibody YL1/2. EMBO. J. 3:12951300. 4. Burns, R. (1987). Tubulins terminal tyrosine. Nature 327:103104.