Cookie Policy: This site uses cookies to improve your experience. You can find out more about our use of cookies in our Privacy Policy. By continuing to browse this site you agree to our use of cookies.
AdipoGen Life Sciences
anti-Polyglutamate chain (polyE), pAb (IN105)
As low as
460
CHF
CHF 460.00
In stock
AG-25B-0030-C05050 µgCHF 460.00

Figure 1: Western blot analysis of protein polyglutamylation by anti-Polyglutamate chain (polyE), pAb (IN105) (Prod. No. AG-25B-0030).
Method: HEK-293T cells are grown in standard culture conditions, transfected with plasmids expressing the tubulin glutamylases TTLL4 and TTLL6 (J. van Dijk, et al.; Mol. Cell 2007) and are lysed in SDS sample buffer and run on a 10% SDS-PAGE. Brain tubulin was prepared following standard procedures (M. Castoldi & A.V. Popov; Protein Expr. Purif. 2003). Different C-terminally modified variants of GST-telokin were produced in bacteria as described before (K. Rogowski, et al.; Cell 2010). The proteins are transferred to a nitrocellulose membrane and detected by standard immunoblot protocol using the anti-polyE, pAb (IN105) (1:2'000) in TBS containing 0.1% Tween-20 for washing steps and 2.5% fat free milk for antibody incubation. The same samples were probed with the anti-Polyglutamylation Modification, mAb (GT335) (Prod. No. AG-20B-0020) for comparison. In the normal HEK-293T cell extracts, no tubulin detection is observed, even on high exposure. Expression of TTLL4 glutamylase does not lead to detection with anti-polyE, pAb (IN105) while glutamylation is observed with GT335. This indicates that TTLL4 generates short glutamate chains on tubulin. Expression of TTLL6 generates long glutamate chains (J. van Dijk, et al.; Mol. Cell 2007), which are detected by anti-polyE, pAb (IN105). Brain tubulin is a heterogenous mixture of different glutamylation states of tubulin (B. Edde, et al.; Science 1990) and therefore strongly detected by anti-polyE, pAb (IN105) and mAb GT335. Anti-polyE, pAb (IN105) also detects gene-encoded linear C-terminal glutamate chains. To illustrate the specificity of anti-polyE, pAb (IN105), recombinant GST-telokin proteins are included in the western blot analysis. Anti-polyE, pAb (IN105) specifically detects glutamate-chains of four and more glutamates. In contrast, GT335 cannot detect such chains (K. Rogowski, et al.; Cell 2010).
Picture courtesy of Dr. Sudarshan Gadadhar & Dr. Carsten Janke, Curie Institute, Paris
Method: HEK-293T cells are grown in standard culture conditions, transfected with plasmids expressing the tubulin glutamylases TTLL4 and TTLL6 (J. van Dijk, et al.; Mol. Cell 2007) and are lysed in SDS sample buffer and run on a 10% SDS-PAGE. Brain tubulin was prepared following standard procedures (M. Castoldi & A.V. Popov; Protein Expr. Purif. 2003). Different C-terminally modified variants of GST-telokin were produced in bacteria as described before (K. Rogowski, et al.; Cell 2010). The proteins are transferred to a nitrocellulose membrane and detected by standard immunoblot protocol using the anti-polyE, pAb (IN105) (1:2'000) in TBS containing 0.1% Tween-20 for washing steps and 2.5% fat free milk for antibody incubation. The same samples were probed with the anti-Polyglutamylation Modification, mAb (GT335) (Prod. No. AG-20B-0020) for comparison. In the normal HEK-293T cell extracts, no tubulin detection is observed, even on high exposure. Expression of TTLL4 glutamylase does not lead to detection with anti-polyE, pAb (IN105) while glutamylation is observed with GT335. This indicates that TTLL4 generates short glutamate chains on tubulin. Expression of TTLL6 generates long glutamate chains (J. van Dijk, et al.; Mol. Cell 2007), which are detected by anti-polyE, pAb (IN105). Brain tubulin is a heterogenous mixture of different glutamylation states of tubulin (B. Edde, et al.; Science 1990) and therefore strongly detected by anti-polyE, pAb (IN105) and mAb GT335. Anti-polyE, pAb (IN105) also detects gene-encoded linear C-terminal glutamate chains. To illustrate the specificity of anti-polyE, pAb (IN105), recombinant GST-telokin proteins are included in the western blot analysis. Anti-polyE, pAb (IN105) specifically detects glutamate-chains of four and more glutamates. In contrast, GT335 cannot detect such chains (K. Rogowski, et al.; Cell 2010).
Picture courtesy of Dr. Sudarshan Gadadhar & Dr. Carsten Janke, Curie Institute, Paris
Product Details | |
---|---|
Product Type | Polyclonal Antibody |
Properties | |
Source/Host | Rabbit |
Immunogen/Antigen | Polyglutamate peptide. |
Application |
Immunocytochemistry: (1:5'000) |
Crossreactivity | All |
Specificity |
Recognizes C-terminally located linear alpha-glutamate chains of 4 and more glutamate residues. |
Purity Detail | Epitope-affinity purified. |
Concentration | Lot dependent |
Formulation | Liquid. In PBS containing 0.02% sodium azide. |
Isotype Negative Control | |
Shipping and Handling | |
Shipping | BLUE ICE |
Short Term Storage | +4°C |
Long Term Storage | +4°C |
Handling Advice | Do not freeze. |
Use/Stability | Stable for at least 1 year after receipt when stored at +4°C. |
Documents | |
MSDS |
![]() |
Product Specification Sheet | |
Datasheet |
![]() |