sepharose

The cell lysates were centrifuged at 14 K rpm, 4°C for 20 min. The supernatant containing recombinant protein was collected and incubated with glutathione sepharose 4B affinity resin (GE Healthcare Bio-Sciences) pre-equilibrated with buffer A.

PLoS ONE Alerts: New Articles Gunjan Arora et al. 2010

For the biofilm inhibition assay, rabbit antibodies were purified using protein G sepharose, according to the manufacturer's instructions (Ab Spin Trap, GE Healthcare) and tested for their ability to inhibit biofilm formation.

PLoS ONE Alerts: New Articles Cira Daniela Rinaudo et al. 2010

The sepharose beads were washed 3 times with 800 µl ice-cold RIPA buffer and resuspended in 60 µl 2× sample buffer, followed by elution through boiling.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

The supernatant was collected in fresh tubes and pre cleared with Protein G sepharose equilibrated with salmon sperm DNA for 30 minutes at 4°C.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

Protein concentration of the dialyzed nuclear extracts was determined by the Bradford method. 30 µg of the dialyzed nuclear extracts were incubated with 2 µg each of anti-Sp1, anti - Sp3 and anti-PPARγ antibodies along with Protein G sepharose beads for 4 hours at 4°C with gentle shaking.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

Nuclear extracts from unstimulated U937 cells were incubated individually with antibodies against Sp1, Sp3 and PPARγ followed by mopping of the immuno-complex with Protein G sepharose beads.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

The cell lysate was pre-cleared by adding 100 µl of 50% Protein G sepharose bead slurry per 1 ml of cell lysate.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

The immune complex was captured by the addition of 100 µl Protein G sepharose.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

The Protein G sepharose beads were washed with cold ChIP sonication buffer containing no protease inhibitors.

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010

The immune complex was precipitated by incubation of the samples with Protein G sepharose equilibrated with salmon sperm DNA at

PLoS ONE Alerts: New Articles Anil K. Singh et al. 2010