Log In Sign up
The largest database of trusted experimental protocols

Reaction buffer

Manufactured by R&D Systems
Visit Supplier

10X reaction buffer is a concentrated solution used to provide the appropriate ionic conditions for various enzymatic reactions. It is typically used as a component in reaction mixtures to maintain the optimal pH and ion concentrations required for the desired enzymatic activity.

Automatically generated - may contain errors

Lab products found in correlation

Sumo1, by R&D Systems (2 mentions)

Spelling variants of this product

E3 Ligase Reaction buffer (7 citations) Ubiquitin Conjugation Reaction Buffer Kit (4 citations) Ubiquitination reaction buffer (3 citations) Ubiquitin conjugation reaction buffer (2 citations) Ubiquitin reaction buffer (2 citations)

2 protocols using reaction buffer

1

SUMO-Mediated MDC1 Ubiquitination

Check if the same lab product or an alternative is used in the 5 most similar protocols
SUMO modification reactions were typically performed in a total volume of 20 μl with 200 ng of SUMO activating enzyme (UBA2) (Boston Biochem), 100 ng of Ubc9 (Boston Biochem), 1 μg of SUMO1 (Boston Biochem), 100 ng of PIAS4 (Novus) and recombinant GST-MDC1 (aa1818–2089) bound to GST-sepharose (20 μg). The 10X reaction buffer (Boston Biochem) was added with 4 mM ATP-Mg. Reactions were incubated at 30°C for 1hr and stopped by addition of E1 stop buffer (Boston Biochem). The beads were washed with NETN and subjected to in vitro ubiquitination assay.
Luo K., Deng M., Li Y., Wu C., Xu Z., Yuan J, & Lou Z. (2015). CDK-mediated RNF4 phosphorylation regulates homologous recombination in S-phase. Nucleic Acids Research, 43(11), 5465-5475.
+ Open protocol
+ Expand
2

In vitro SUMOylation Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro SUMOylation assays were performed as previously described with some changes61 (link). In brief, SUMO modification reactions were typically performed in a total volume of 100 μl with 200 ng of SUMO-activating enzyme (SAE1) (Boston Biochem), 100 ng of UBC9 (Boston Biochem), 1 μg of SUMO1 (Boston Biochem), 100 ng of PIAS2 (NOVUS), and recombinant GST-HNRNPA2 bound to GST-sepharose (20 μg). The 10X reaction buffer (Boston Biochem) was added with 5 mM ATP-Mg. Reactions were incubated at 37°C for 1h and stopped by adding 10 mM EDTA. The beads were washed with NETN buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40 with protease inhibitors) for 3 times and boiled in 1X LDS loading buffer. Finally, the samples were analyzed by immunoblotting.
Zhu S., Hou J., Gao H., Hu Q., Kloeber J.A., Huang J., Zhao F., Zhou Q., Luo K., Wu Z., Tu X., Yin P, & Lou Z. (2023). SUMOylation of HNRNPA2B1 modulates RPA dynamics during unperturbed replication and genotoxic stress responses. Molecular cell, 83(4), 539-555.e7.
+ 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!