Analytical gel filtrations of recombinant NefSF2, SH3B6, sdAb19, Neffin, and all mutants thereof were performed using a multicomponent Waters 626 LC system (Waters, MA) equipped with a Superdex S75 (10/300 GL) column (GE Healthcare). Typically, 100 μl of a 1.5 mg/ml protein solution was loaded onto the column that was equilibrated in 10 mM Tris/HCl (pH 9.0), 100 mM NaCl buffer prior to injection of the protein samples. Gel filtrations were run at a flow rate of 0.5 ml per minute in 10 mM Tris/HCl (pH 9.0), 100 mM NaCl onto the S75 column at 4°C or 20°C. The optical density was monitored at a wavelength of 280 nm over the time course of the experiment. Gel filtration experiments were performed repeated times.
626 lc system
Manufactured by Waters Corporation
Sourced in United States
The 626 LC System is a liquid chromatography instrument designed for laboratory analysis. It provides reliable separation and detection of chemical compounds in liquid samples. The system features precise solvent delivery, automated sample handling, and advanced data processing capabilities to support a range of analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using 626 lc system
1
Analytical Gel Filtration of Recombinant Proteins
2
Histones Fractionation by RP-HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histones were fractionated by RP-HPLC according to the method described by Shechter et al., 2007 (link). After lyophilisation samples were resuspended in 300 µl Buffer A (5% acetonitrile, 0.1% trifluoroacetic acid) and centrifuged at 14,000 g to remove particulate matter. 150 µl of clarified sample was mixed with 40 µl Buffer A before being applied to a C18 column (#218TP53, Grave Vydac) and subjected to RP-HPLC (Waters 626 LC System, MA, US) as described by Shechter et al., 2007 (link). The flow rate was set to 1 ml min–1 and fractions were collected at 30 s intervals from minute 30–55. All fractions were lyophilised and stored at –80 °C until analysis. To determine which fractions contained H3 and cleaved species, each fraction was dissolved in 50 µl dH2O and 5 µl was subjected to SDS-PAGE and either stained with Coomassie blue stain or transferred to PVDF and immunoblotted for H3 and H4 as described already.
3
RP-HPLC Fractionation of Histones
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histones were fractionated by RP-HPLC according to the method described by Shechter et al (2007) (link). After lyophilisation samples were resuspended in 300 µl Buffer A (5% acetonitrile, 0.1% trifluoroacetic acid) and centrifuged at 14,000 g to remove particulate matter. 150 µl of clarified sample was mixed with 40 µl Buffer A before being applied to a C18 column (#218TP53, Grave Vydac) and subjected to RP-HPLC (Waters 626 LC System, MA, US) as described by Schecter et al (2007) . The flow rate was set to 1 ml min -1 and fractions were collected at 30 s intervals from minute 30 to 55. All fractions were lyophilised and stored at -80 °C until analysis. To determine which fractions contained H3 and cleaved species, each fraction was dissolved in 50 µl dH2O and 5 µl was subjected to SDS-PAGE and either stained with Coomassie blue stain or transferred to PVDF and immunoblotted for H3 and H4 as described already.
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
Revolutionizing how scientists
search and build protocols!