RIPA Buffer (10X)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Whole cell extracts were prepared by lysing the cells with ice-cold RIPA buffer following the manufacturer's protocol.

Protocol tips
- Whole cell extracts were prepared by lysing the cells with ice-cold RIPA buffer following the manufacturer's protocol.

Publication protocol

Primary human gingival cells (HGEPs) were purchased from CellnTec and maintained according to the manufacturer’s instructions. One µg/mL of Escherichia coli (O55:B5) lipopolysaccharide (LPS; Sigma-Aldrich) was added to stimulate HGEPs for 0, 4, 8, and 24 h. Whole cell extracts were prepared by lysing the cells with ice-cold RIPA buffer (Cell Signaling 10× buffer and Roche protease inhibitor mixture). The protein concentrations of the lysates were determined by DC Protein Assay (Bio-Rad). The same amount of protein was separated by 4% to 12% Bis-Tris gels and transferred to a nitrocellulose membrane (BD). The membrane was blocked 1 h in 5% milk powder–TBST solution and then overnight blotted with anti-G6PD (H160, Santa Cruz) or anti-β actin antibodies (13E5, Cell Signaling). Membranes were washed 3 times with TBST, incubated with anti-rabbit HRP-labeled secondary antibodies in TBST, and developed with the ECL system (SuperSignal West Pico; Pierce).

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 isolation Mammalian cells - Human gingival epithelial cells using RIPA Buffer (10X) from Cell Signaling Technology.

Manufacturer protocol

Download the product protocol from Cell Signaling Technology for RIPA Buffer (10X) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Protein isolation Mammalian cells - Human gingival epithelial cells using RIPA Buffer (10X) from Cell Signaling Technology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.