Log In Sign up
The largest database of trusted experimental protocols

Recombinant ubiquitin

Manufactured by R&D Systems
Visit Supplier

Recombinant ubiquitin is a purified protein produced using recombinant DNA technology. Ubiquitin is a small regulatory protein that plays a role in cellular processes such as protein degradation and modification.

Automatically generated - may contain errors

Lab products found in correlation

Ubch5b, by R&D Systems (2 mentions) Adenosine triphosphate (atp), by Merck Group (2 mentions) Ubiquitin vinyl sulfone, by R&D Systems (1 mentions) Nms 873, by Apexbio (1 mentions)

5 protocols using recombinant ubiquitin

1

Xenopus Egg Extract Plasmid Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Xenopus egg extracts were prepared as described previously75 . Plasmid DNA was incubated in nucleoplasmic extract (NPE) at a final concentration of 50 ng/μL at 21 °C. Plasmids used in this study do not contain DNA damage and are not replicated during the reaction due to the absence of licensing in a high-speed supernatant (HSS) extract46 (link). Where indicated, reactions were supplemented with: 150 mM NMS-873 (ApexBio Technology), 500 mM CB-5083 (PharmaBlock Sciences), 150 μM Ubiquitin Vinyl Sulfone (Boston Biochem), 50 μM recombinant ubiquitin (Boston Biochem), or 40 ng/μL recombinant p97. All experiments were performed three or more times and representative results are shown. Experiments involving vertebrate animals (Xenopus laevis) were performed in accordance with all guidelines and regulations using an approved protocol administered by the Medical University of South Carolina Institutional Animal Care and Use Committee (IACUC).
Rycenga H.B., Wolfe K.B., Yeh E.S, & Long D.T. (2019). Uncoupling of p97 ATPase activity has a dominant negative effect on protein extraction. Scientific Reports, 9, 10329.
+ Open protocol
+ Expand
2

CARD9, BCL10, and CARD11 Protein Study

Check if the same lab product or an alternative is used in the 5 most similar protocols
The antibodies used in this study were listed in Supplementary Table S3. Recombinant Ubiquitin and derivatives were from Boston Biochem Inc. (Cambridge, MA). Recombinant CARD9 (ab131800) and BCL10 (ab82241) were obtained from Abcam Inc. (Cambridge, MA). Recombinant CARD11 was obtained from Creative BioMart (Shirley, NY; Card11-385M). Other non-specified reagents were purchased from Sigma-Aldrich (St. Louis, MO).
Yu Z., Li X., Yang M., Huang J., Fang Q., Jia J., Li Z., Gu Y., Chen T, & Cao X. (2021). TRIM41 is required to innate antiviral response by polyubiquitinating BCL10 and recruiting NEMO. Signal Transduction and Targeted Therapy, 6, 90.
+ Open protocol
+ Expand
3

In vitro Ubiquitination Assay of Arc Protein

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro ubiquitination of Arc was performed using 0.1 μg of recombinant ubiquitin E1 (UBE1) (Boston Biochem), 0.5 μg recombinant ubiquitin E2 (UbcH7) (Boston Biochem), 10 μg of recombinant ubiquitin (Boston Biochem), and 0.1 μg immunopurified Flag-Triad3A added to 0.25 μg of His-tagged Arc (or 0.15 μg of GST-Arc full-length or GST-Arc 132–396) in the presence of an ATP regenerating system (10 mM creatine phosphate, 10 units of creatine kinase, 1 unit inorganic pyrophosphatase, and 2 mM ATP) in 50 mM Tris (pH 7.6) and 5 mM MgCl2 in a 120 μl reaction with 1 μg/mL aprotinin and 1 mM PMSF. GST-Arc was purchased from BD Pharmingen. GST-Arc (132–396) was expressed and purified as previously described (Chowdhury et al., 2006 (link); Greer et al., 2010 (link)). The reaction was incubated at 30°C for 120 min. Reactions were terminated by boiling samples in SDS-PAGE loading buffer, polypeptides resolved by SDS-PAGE gel, and immunoblot analysis performed with an anti-His antibody to detect ubiquitinated bands or with anti-Flag antibody to detect immunopurified Flag-Triad3A. For additional assay information, see supplemental methods.
Mabb A.M., Je H.S., Wall M.J., Robinson C.G., Larsen R.S., Qiang Y., Corrêa S.A, & Ehlers M.D. (2014). Triad3A Regulates Synaptic Strength by Ubiquitination of Arc. Neuron, 82(6), 1299-1316.
+ Open protocol
+ Expand
4

MERS-CoV ORF3 Ubiquitination by HUWE1

Check if the same lab product or an alternative is used in the 5 most similar protocols
The MERS-CoV ORF3 and the HUWE1 HECT domain of WT and C4341A mutant were cloned and expressed as GST-fusion proteins. All the proteins were expressed in BL21 bacterial cells and purified with Glutathione Sepharose beads. The beads with ORF3 protein were incubated with HUWE1 HECT WT or C4341A mutant, together with 200 ng of E1 (UBE1, R&D system), 400 ng of E2 (UbcH5b, R&D system), and 2 μg of recombinant ubiquitin (R&D system) in reaction buffer (50 mM Tris [PH 7.4], 2 mM MgCl2, 4 mM ATP [Sigma]) at 37 °C for 1 h. The supernatant was removed, and the reaction was terminated by adding 2x protein loading buffer. The ubiquitin chain was detected by immunoblotting.
Zhou Y., Zheng R., Liu S., Disoma C., Du A., Li S., Chen Z., Dong Z., Zhang Y., Li S., Liu P., Razzaq A., Chen X., Liao Y., Tao S., Liu Y., Xu L., Zhang Q., Peng J., Deng X., Li S., Jiang T, & Xia Z. (2022). Host E3 ligase HUWE1 attenuates the proapoptotic activity of the MERS-CoV accessory protein ORF3 by promoting its ubiquitin-dependent degradation. The Journal of Biological Chemistry, 298(2), 101584.
+ Open protocol
+ Expand
5

Ubiquitination of MERS-CoV ORF4b by UBR5

Check if the same lab product or an alternative is used in the 5 most similar protocols
The MERS-CoV ORF4b was cloned and expressed as GST fusion proteins. The target viral protein was expressed in BL21 bacterial cells and purified with glutathione Sepharose beads. In addition, the Flag-UBR5 and Flag-UBR5-C2768A were purified from HEK293T cells. The beads with ORF4b protein were incubated with UBR5-WT or C2768A mutant, together with 200 ng of E1 (UBE1; R&D Systems), 400 ng of E2 (UbcH5b; R&D Systems), and 2 μg of recombinant ubiquitin (R&D Systems) in reaction buffer (50 mM Tris [PH 7.4], 2 mM MgCl2, 4 mM ATP [Sigma]) at 37°C for 1 h. The supernatant was removed, and the reaction was terminated by adding 2× protein loading buffer. The ubiquitin chain was detected by immunoblotting.
Zhou Y., Zheng R., Liu D., Liu S., Disoma C., Li S., Liao Y., Chen Z., Du A., Dong Z., Zhang Y., Liu P., Razzaq A., Chen D., Chen X., Zhong X., Liu S., Tao S., Liu Y., Xu L., Deng X., Li J., Jiang T., Zhao J., Li S, & Xia Z. (2022). UBR5 Acts as an Antiviral Host Factor against MERS-CoV via Promoting Ubiquitination and Degradation of ORF4b. Journal of Virology, 96(17), e00741-22.
+ 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!