AdipoGen Life Sciences

anti-beta-Tubulin (beta-monoE) pAb (IN115)

Product Code:
 
AG-25B-0039
Product Group:
 
Primary Antibodies
Host Type:
 
Rabbit
Antibody Clonality:
 
Polyclonal
Regulatory Status:
 
RUO
Target Species:
  • Human
  • Mouse
Applications:
  • Immunocytochemistry (ICC)
  • Immunohistochemistry (IHC)
  • Immunoprecipitation (IP)
  • Western Blot (WB)
Shipping:
 
BLUE ICE
Storage:
 
Short Term Storage: +4?C. Long Term Storage: -20?C
1 / 1
Figure 1: Immunoblot analysis of protein glutamylation with anti-?-Tubulin (?-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 ?- and ?-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 ?-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 ?-tubulin is detected with ?-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 ?-monoE detects only ?-tubulin, while expression of TTLL7 generates ?-monoE-positive ?- and ?-tubulin. In the purified brain tubulin, the antibody specifically detects glutamylation of ?-tubulin. Picture courtesy of Sudarshan Gadadhar, Shreyangi Chakraborty, Mariya Genova and Carsten Janke, Institut Curie, Orsay.
❮❯

Figure 1: Immunoblot analysis of protein glutamylation with anti-?-Tubulin (?-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 ?- and ?-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 ?-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 ?-tubulin is detected with ?-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 ?-monoE detects only ?-tubulin, while expression of TTLL7 generates ?-monoE-positive ?- and ?-tubulin. In the purified brain tubulin, the antibody specifically detects glutamylation of ?-tubulin. Picture courtesy of Sudarshan Gadadhar, Shreyangi Chakraborty, Mariya Genova and Carsten Janke, Institut Curie, Orsay.

No additional charges, what you see is what you pay! *

CodeSizePrice
AG-25B-0039-C05050 ug£350.00
Quantity:
Prices exclude any Taxes / VAT
How to order with us   Contact us
Stay in control of your spending. These prices have no additional charges, not even shipping!
* Rare exceptions are clearly labelled (only 0.14% of items!).
Multibuy discounts available! Contact us to find what you can save.
This product comes from: Switzerland.
Typical lead time: 7-10 working days.
Contact us for more accurate information.
  • Further Information
  • Documents
  • References
  • Show All

Further Information

Concentration:
1mg/ml
EClass:
32160000
Form (Short):
liquid
Formulation:
Liquid. In PBS containing 0.02% Proclin 300.
Handling Advice:
After opening, prepare aliquots and store at -20°C.Avoid freeze/thaw cycles.
Immunogen:
Synthetic peptide corresponding to D431EQGEFE(E-COOH*)EEEG441-NH2 of human tubulin beta-2A chain (*secondary glutamate branched from gamma-carboxyl group of glutamate as isopeptide bond).
Long Description:
Polyclonal Antibody. Recognizes the posttranslational modification glutamylation with specific reactivity for the GE(*-E)F motive. Because the sequence GEF is specific to several beta-tubulin isotypes, the antibody selectively detects mono-glutamylated beta-tubulin. Applications: ICC, IHC, IP, WB. Source: Rabbit. 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, alpha- and beta-tubulin. It is catalyzed by multiple enzymes that belong to the family of tubulin tyrosine-ligase like (TTLL) enzymes. These TTLLs have substrate (alpha- vs beta-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 beta-monoE antibody selectively labels glutamylation of beta-tubulin due to its specificity to a sequence motif (GE(*-E)F) that only exists in beta-tubulin isotypes. It allows to specifically detect beta-tubulin glutamylation in brain tubulin, where other antibodies predominantly detect alpha-tubulin glutamylation.
NCBI, Uniprot Number:
Q13885
Other data:
Recognizes predominantly mono-glutamylated beta-tubulin. However, in cells overexpressing enzymes that can modify both alpha- and beta-tubulin, it might detect glutamylation on alpha-tubulin as well. Requires GE(*-E)F motive, which is present in several beta-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.
Package Type:
Plastic Vial
Product 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, alpha- and beta-tubulin. It is catalyzed by multiple enzymes that belong to the family of tubulin tyrosine-ligase like (TTLL) enzymes. These TTLLs have substrate (alpha- vs beta-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 beta-monoE antibody selectively labels glutamylation of beta-tubulin due to its specificity to a sequence motif (GE(*-E)F) that only exists in beta-tubulin isotypes. It allows to specifically detect beta-tubulin glutamylation in brain tubulin, where other antibodies predominantly detect alpha-tubulin glutamylation.
Purity:
>95% (SDS-PAGE)
Source / Host:
Rabbit
Specificity:
Recognizes the posttranslational modification glutamylation with specific reactivity for the GE(*-E)F motive. Because the sequence GEF is specific to several beta-tubulin isotypes, the antibody selectively detects mono-glutamylated beta-tubulin.
Transportation:
Non-hazardous
UNSPSC Category:
Primary Antibodies
UNSPSC Number:
12352203
Use & Stability:
Stable for at least 1 year after receipt when stored at -20°C.

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)