Edc4 ge 1 rabbit antibody

A solution containing 5% (wt/v%) bovine serum albumin dissolved in PBS was added to cells and incubated for 1 h at RT while shaking. The solution was aspirated and a new solution containing 5% (wt/v%) bovine serum albumin and EDC4/GE-1 rabbit antibody (Cell Signaling Technology, cat#2548S; 1:400) dissolved in PBS was incubated with the cells at 4 °C overnight with shaking. The cells were washed and a solution containing Alexa Fluor^® 488 Conjugate Anti-Rabbit IgG (H+L) F(ab’)2 Fragment (Cell Signaling Technology, cat#4412S; 1:1,000) and DAPI (Tocris, 1:2,000) dissolved in PBST was incubated with cells (1 mL) at 37 °C with 5% CO₂ for 1 h. The cells were washed and mounted as described in the above protocols.

The same protocol was followed as stated above for EDC4 except the antibody solutions and times used were as follows: primary antibody solution: 5% (wt/v%) bovine serum albumin, DYKDDDDK Tag mouse mAb, (Cell Signaling Technology, cat#8146, 1:1,000), and EDC4/GE-1 rabbit antibody (Cell Signaling Technology, cat#2548S; 1:400) dissolved in PBS were added to wells (1 mL) and cells were shaken overnight at 4°C; secondary antibody solution: Alexa Fluor^® 488 Conjugate Anti-Rabbit IgG (H+L) F(ab’)2 Fragment (Cell Signaling Technology, cat#4412S; 1:1,000), Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 650 (Invitrogen, cat#84545, 1:500) and DAPI (Tocris, 1:2,000) dissolved in PBST was incubated with cells (1 mL) at 37°C with 5% CO₂ for 1 hour.