SRAPd was incubated with the abasic site containing 3’ overhang DNA (same as used in SRAPd_DPC co-crystallization) at 1:1.2 ratio in a buffer containing 150 mM NaCl, 20 mM Hepes pH 7.5, 10 mM MgCl2 at room temperature overnight. All LC-MS data were acquired according to the previously published protocol23 , on an Agilent 6545 Q-TOF (Santa Clara, CA) equipped with a Dual Agilent Jet Stream ESI source coupled with an Agilent 1260 Infinity HPLC system (Santa Clara, CA). The analytical column utilized was a 300 StableBond Poroshell (Agilent, part number 883750–909) 2.1 × 100-mm-i.d. reversed-phase C3 (5 μm particle size). Mobile phase (A) consisted of 97% HPLC grade water with 0.5% formic acid and 2.5% ACN, while mobile phase (B) was 96% ACN with 0.5% formic acid and 3.5% HPLC grade water. A gradient profile was utilized at a flow rate of 500 μL/min. The mobile phase was held for 2 min at 5% B (with eluant going to waste) and then switched to the mass spectrometer from 2–6 min during which time solvent B increased from 5–95%. Two microliters of a 30μM solution of each sample was injected. Raw data files were analyzed by Agilent MassHunter BioConfirm software (vB.07.00). Mass spectra between 4 and 6 minutes were extracted, averaged and deconvoluted using the MaxEnt algorithm.
300 stablebond poroshell
Manufactured by Agilent Technologies
The 300 StableBond Poroshell is a high-performance liquid chromatography (HPLC) column produced by Agilent Technologies. It is designed to provide stable and consistent separation performance for a wide range of analytical applications.
Automatically generated - may contain errors
2 protocols using 300 stablebond poroshell
1
Characterization of SRAPd-Abasic DNA Complex
2
Characterization of SRAPd-Abasic DNA Complex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SRAPd was incubated with the abasic site containing 3’ overhang DNA (same as used in SRAPd_DPC co-crystallization) at 1:1.2 ratio in a buffer containing 150 mM NaCl, 20 mM Hepes pH 7.5, 10 mM MgCl2 at room temperature overnight. All LC-MS data were acquired according to the previously published protocol23 , on an Agilent 6545 Q-TOF (Santa Clara, CA) equipped with a Dual Agilent Jet Stream ESI source coupled with an Agilent 1260 Infinity HPLC system (Santa Clara, CA). The analytical column utilized was a 300 StableBond Poroshell (Agilent, part number 883750–909) 2.1 × 100-mm-i.d. reversed-phase C3 (5 μm particle size). Mobile phase (A) consisted of 97% HPLC grade water with 0.5% formic acid and 2.5% ACN, while mobile phase (B) was 96% ACN with 0.5% formic acid and 3.5% HPLC grade water. A gradient profile was utilized at a flow rate of 500 μL/min. The mobile phase was held for 2 min at 5% B (with eluant going to waste) and then switched to the mass spectrometer from 2–6 min during which time solvent B increased from 5–95%. Two microliters of a 30μM solution of each sample was injected. Raw data files were analyzed by Agilent MassHunter BioConfirm software (vB.07.00). Mass spectra between 4 and 6 minutes were extracted, averaged and deconvoluted using the MaxEnt algorithm.
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!