Antibody Type: Recombinant Rabbit Monoclonal

Application: WB,ICC/IF,FCM

Reactivity: Human

IgG

7Z785

Human

1. Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-FOXP3 antibody (TMAB-00711)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. 25 μg total protein per Lane of various lysates probed with FOXP3 monoclonal antibody, unconjugated (TMAB-00711) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.
3. The MJ (H) cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C,the cells then were incubated in 5% BSA to block non-specific protein-protein interactions (30 min at room temperature). Primary Antibody (green): Rabbit Anti-FOXP3 antibody (TMAB-00711,1:100); Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488: 1 μg/test. Isotype Control (orange): Rabbit IgG. Blank control (black): PBS.
4. 4% Paraformaldehyde-fixed MJ (H) cell; Triton X-100 at RT for 20 min; Antibody incubation with (FOXP3) monoclonal Antibody, unconjugated (TMAB-00711) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green) at 37°C for 90 min, DAPI (blue) was used to stain the cell nucleus. PBS instead of the primary antibody was used as the blank control.

WB,ICC/IF,FCM

WB: 1:500-2000; ICC/IF: 1:50-200; FCM: 1:50-100

Monoclonal

Recombinant Protein: human FOXP3 protein, around C-terminal 150aa

Human

50943

Chromatid Cohesion,FOXP,Regulatory T Cells