pBud-hIL-7

Protocol tips

Upstream tips	Protocol tips	Downstream tips
The CHO-K1 cells were passaged 24 h before the transfections and harvested at 75-85% confluency on the day of transfection.	For stable transfections, 5×105 cells and 20 µg of each plasmid (either circular plasmid or plasmid linearized with XhoI) were used at 350 V and 950 µF in a Gene Pulser Xcell electroporation system (Bio-Rad, CA, USA) using a single exponentially-decaying pulse. The electroporated cells were recovered in a shaking flask containing growth medium.	At 48 h after electroporation, cell viability was measured by trypan blue exclusion on a hemocytometer, and supernatants were collected for IL-7 expression analyses using the enzyme-linked immunosorbent assay (ELISA). Transfected cells were cultured with complete medium supplemented with 600 µg/ml of Zeocin. Pools of stable clones were assayed after 14 days of selection. The amount of protein in the supernatant was assessed by the Bradford protein assay (20).

Upstream tips
The CHO-K1 cells were passaged 24 h before the transfections and harvested at 75-85% confluency on the day of transfection.
Protocol tips
For stable transfections, 5×105 cells and 20 µg of each plasmid (either circular plasmid or plasmid linearized with XhoI) were used at 350 V and 950 µF in a Gene Pulser Xcell electroporation system (Bio-Rad, CA, USA) using a single exponentially-decaying pulse. The electroporated cells were recovered in a shaking flask containing growth medium.
Downstream tips
At 48 h after electroporation, cell viability was measured by trypan blue exclusion on a hemocytometer, and supernatants were collected for IL-7 expression analyses using the enzyme-linked immunosorbent assay (ELISA). Transfected cells were cultured with complete medium supplemented with 600 µg/ml of Zeocin. Pools of stable clones were assayed after 14 days of selection. The amount of protein in the supernatant was assessed by the Bradford protein assay (20).

Publication protocol

The CHO-K1 cells were passaged 24 h before the transfections and harvested at 75-85% confluency on the day of transfection. For stable transfections, 5×105 cells and 20 µg of each plasmid (either circular plasmid or plasmid linearized with XhoI) were used at 350 V and 950 µF in a Gene Pulser Xcell electroporation system (Bio-Rad, CA, USA) using a single exponentially-decaying pulse. The electroporated cells were recovered in a shaking flask containing growth medium. At 48 h after electroporation, cell viability was measured by trypan blue exclusion on a hemocytometer, and supernatants were collected for IL-7 expression analyses using the enzyme-linked immunosorbent assay (ELISA). Transfected cells were cultured with complete medium supplemented with 600 µg/ml of Zeocin. Pools of stable clones were assayed after 14 days of selection. The amount of protein in the supernatant was assessed by the Bradford protein assay (20).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Eukaryotic cells - CHO hIL-7 using pBud-hIL-7 from Abbas Ghaderi, Shiraz Institute for Cancer Research, School of M.

Manufacturer protocol

Download the product protocol from Abbas Ghaderi, Shiraz Institute for Cancer Research, School of M for pBud-hIL-7 below.

Videos

Check out videos that might be relevant for performing Protein Expression Eukaryotic cells - CHO hIL-7 using pBud-hIL-7 from Abbas Ghaderi, Shiraz Institute for Cancer Research, School of M. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.