Log In Sign up
The largest database of trusted experimental protocols

E2 ubiquitin conjugating enzyme ubch7

Manufactured by R&D Systems
Visit Supplier

E2 ubiquitin conjugating enzyme (UbcH7) is a core component of the ubiquitin-proteasome system. It functions as an ubiquitin-conjugating enzyme, facilitating the transfer of ubiquitin from E1 ubiquitin-activating enzyme to target proteins, marking them for proteasomal degradation.

Automatically generated - may contain errors

Lab products found in correlation

Anti v5 agarose beads, by Merck Group (2 mentions) Ubiquitin activating enzyme e1, by R&D Systems (2 mentions) Ubiquitin, by Merck Group (2 mentions) Protease inhibitor, by Roche (2 mentions)

Close spelling variants of this product

E2 (UbcH7; E2 enzyme UbcH7 Ubiquitin

2 protocols using e2 ubiquitin conjugating enzyme ubch7

1

Detecting Ubiquitination of NEDD4L Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunoprecipitation assay, transfected cells were lysed with RIPA buffer supplemented with protease inhibitors (Roche), MG-132 (25 µM) and PR-619 (20mM) and protein extracts were incubated with anti-V5 agarose beads (A7345, Sigma) for 2 hours at 4°C under constant rotation in RIPA buffer. Immunoprecipitated proteins were eluted in Laemmli SDS buffer at 95°C and then subjected to SDS-PAGE. For in vitro ubiquitination assay, immunopurified NEDD4L from transfected N2A cells was incubated in reaction mixtures containing 200nM E1 ubiquitin-activating enzyme (BostonBiochem, Cambridge, MA), 400 nM E2 ubiquitin conjugating enzyme (UbcH7; BostonBiochem), 400 μM of ubiquitin (Sigma) and 2 mM ATP in a reaction buffer (25mM Tris/HCl (pH 7.5), 50 mM NaCl, 0.1 μM dithiothreitol and 4 mM MgCl2). Reactions were incubated for 1hour at 30°C and analyzed by immunoblotting with anti-ubiquitin, anti-V5 and anti-NEDD4L antibodies.
Broix L., Jagline H., Ivanova E., Schmucker S., Drouot N., Clayton-Smith J., Pagnamenta A.T., Metcalfe K.A., Isidor B., Louvier U.W., Poduri A., Taylor J.C., Tilly P., Poirier K., Saillour Y., Lebrun N., Stemmelen T., Rudolf G., Muraca G., Saintpierre B., Elmorjani A., Moïse M., Weirauch N.B., Guerrini R., Boland A., Olaso R., Masson C., Tripathy R., Keays D., Beldjord C., Nguyen L., Godin J., Kini U., Nischké P., Deleuze J.F., Bahi-Buisson N., Sumara I., Hinckelmann M.V, & Chelly J. (2016). Mutations in the HECT domain of NEDD4L lead to AKT/mTOR pathway deregulation and cause periventricular nodular heterotopia. Nature genetics, 48(11), 1349-1358.
+ Open protocol
+ Expand
2

Detecting Ubiquitination of NEDD4L Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunoprecipitation assay, transfected cells were lysed with RIPA buffer supplemented with protease inhibitors (Roche), MG-132 (25 µM) and PR-619 (20mM) and protein extracts were incubated with anti-V5 agarose beads (A7345, Sigma) for 2 hours at 4°C under constant rotation in RIPA buffer. Immunoprecipitated proteins were eluted in Laemmli SDS buffer at 95°C and then subjected to SDS-PAGE. For in vitro ubiquitination assay, immunopurified NEDD4L from transfected N2A cells was incubated in reaction mixtures containing 200nM E1 ubiquitin-activating enzyme (BostonBiochem, Cambridge, MA), 400 nM E2 ubiquitin conjugating enzyme (UbcH7; BostonBiochem), 400 μM of ubiquitin (Sigma) and 2 mM ATP in a reaction buffer (25mM Tris/HCl (pH 7.5), 50 mM NaCl, 0.1 μM dithiothreitol and 4 mM MgCl2). Reactions were incubated for 1hour at 30°C and analyzed by immunoblotting with anti-ubiquitin, anti-V5 and anti-NEDD4L antibodies.
Broix L., Jagline H., Ivanova E., Schmucker S., Drouot N., Clayton-Smith J., Pagnamenta A.T., Metcalfe K.A., Isidor B., Louvier U.W., Poduri A., Taylor J.C., Tilly P., Poirier K., Saillour Y., Lebrun N., Stemmelen T., Rudolf G., Muraca G., Saintpierre B., Elmorjani A., Moïse M., Weirauch N.B., Guerrini R., Boland A., Olaso R., Masson C., Tripathy R., Keays D., Beldjord C., Nguyen L., Godin J., Kini U., Nischké P., Deleuze J.F., Bahi-Buisson N., Sumara I., Hinckelmann M.V, & Chelly J. (2016). Mutations in the HECT domain of NEDD4L lead to AKT/mTOR pathway deregulation and cause periventricular nodular heterotopia. Nature genetics, 48(11), 1349-1358.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!