Log In Sign up
The largest database of trusted experimental protocols

E1 uba1

Manufactured by R&D Systems
Visit Supplier

The E1 (UBA1) is a lab equipment product offered by R&D Systems. Its core function is to activate the ubiquitin-activating enzyme E1, which is a critical step in the ubiquitin-proteasome system.

Automatically generated - may contain errors

Lab products found in correlation

E2 ubch5a, by R&D Systems (3 mentions) Amersham 600 ccd based imaging system, by GE Healthcare (2 mentions)

3 protocols using e1 uba1

1

In vitro ubiquitination of CDK12-cycK

Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro ubiquitination was performed by mixing N8DDB1-CUL4-RBX1 at 70 nM with a reaction mixture containing kinase inhibitors at 2 μM, CDK12-cycK at 500 nM, E1 (UBA1, BostonBiochem) at 50 nM, E2 (UBCH5a, BostonBiochem) at 1 μM, and ubiquitin at 20 μM. Reactions were carried out in 50 mM Tris (pH 7.5), 150 mM NaCl, 5 mM MgCl2, 0.2 mM CaCl2, 1 mM ATP, 0.1% Triton X-100 and 0.1 mg/mL BSA, incubated for 0–30 minutes at 30°C and analysed by western blot using anti-cycK and anti-rabbit IgG antibodies. Blots were scanned on an Amersham 600 CCD-based imaging system (GE Life Sciences).
Słabicki M., Kozicka Z., Petzold G., Li Y.D., Manojkumar M., Bunker R., Donovan K.A., Sievers Q.L., Koeppel J., Suchyta D., Sperling A.S., Fink E.C., Gasser J.A., Wang L.R., Corsello S.M., Sellar R.S., Jan M., Gillingham D., Scholl C., Fröhling S., Golub T.R., Fischer E.S., Thomä N.H, & Ebert B.L. (2020). The CDK inhibitor CR8 acts as a molecular glue degrader that depletes cyclin K. Nature, 585(7824), 293-297.
+ Open protocol
+ Expand
2

In vitro Ubiquitination of ZFP91

Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro ubiquitination was performed by mixing ZFP91 at 0.6 μM, and N8CRL4CRBN at 80 nM with a reaction mixture containing IMiDs at indicated concentrations or a DMSO control, E1 (UBA1, Boston Biochem) at 40 nM, E2 (UBCH5a, Boston Biochem) at 1.2 μM, ubiquitin at 23 μM. Reactions were carried out in 50 mM Tris pH 7.5, 30 mM NaCl, 5 mM MgCl2, 0.2 mM CaCl2, 2.5 mM ATP, 0.1% Triton X-100 and 0.1 mg ml−1 BSA, incubated for 15–30 min at 30 °C and analysed by western blot using anti-ZFP91 and anti-rabbit IRDye 800CW antibodies. Blots were scanned on a LICOR Odyssey infrared imaging system (uncropped immunoblots are provided in Supplementary Fig. 4).
An J., Ponthier C.M., Sack R., Seebacher J., Stadler M.B., Donovan K.A, & Fischer E.S. (2017). pSILAC mass spectrometry reveals ZFP91 as IMiD-dependent substrate of the CRL4CRBN ubiquitin ligase. Nature Communications, 8, 15398.
+ Open protocol
+ Expand
3

In vitro ubiquitination of CDK12-cycK

Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro ubiquitination was performed by mixing N8DDB1-CUL4-RBX1 at 70 nM with a reaction mixture containing kinase inhibitors at 2 μM, CDK12-cycK at 500 nM, E1 (UBA1, BostonBiochem) at 50 nM, E2 (UBCH5a, BostonBiochem) at 1 μM, and ubiquitin at 20 μM. Reactions were carried out in 50 mM Tris (pH 7.5), 150 mM NaCl, 5 mM MgCl2, 0.2 mM CaCl2, 1 mM ATP, 0.1% Triton X-100 and 0.1 mg/mL BSA, incubated for 0–30 minutes at 30°C and analysed by western blot using anti-cycK and anti-rabbit IgG antibodies. Blots were scanned on an Amersham 600 CCD-based imaging system (GE Life Sciences).
Słabicki M., Kozicka Z., Petzold G., Li Y.D., Manojkumar M., Bunker R., Donovan K.A., Sievers Q.L., Koeppel J., Suchyta D., Sperling A.S., Fink E.C., Gasser J.A., Wang L.R., Corsello S.M., Sellar R.S., Jan M., Gillingham D., Scholl C., Fröhling S., Golub T.R., Fischer E.S., Thomä N.H, & Ebert B.L. (2020). The CDK inhibitor CR8 acts as a molecular glue degrader that depletes cyclin K. Nature, 585(7824), 293-297.
+ 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!