Antibody Type: Rabbit Polyclonal

Application: ICC/IF,FCM

Reactivity: Human,Mouse

IgG

Human,Mouse

1. Blank control: THP-1. Primary Antibody (green line): Rabbit Anti-TNF alpha antibody (TMAB-01871)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Protocol
The cells were treated with TPA (80 nM, overnight) and then treated with LPS (1 μg/mL, 18 hr/6 hr) and Brefeldin A (300 ng/mL, last 3 hr of stimulation). The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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. Blank control: Raw264.7. Primary Antibody (green line): Rabbit Anti-TNF alpha antibody (TMAB-01871)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Protocol
The cells were treated with LPS (1 μg/mL, 18 hr/6 hr) and Brefeldin A (300 ng/mL, last 3 hr of stimulation). The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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.
3. 4% Paraformaldehyde-fixed THP-1 (LPS treated (100 ng/ml, 16 h) and Brefeldin A treated (5 µg/ml, 4 h)) (H) cell; Antibody incubation with (TNF alpha) polyclonal Antibody, unconjugated (TMAB-01871) 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.

ICC/IF,FCM

ICC/IF: 1:50-200; FCM: 1ug/Test

Polyclonal

Recombinant Protein: human TNF alpha protein

Human

7124

TNF,Inflammatory mediators,TNF Superfamily,Metabolic disorders,SARS Coronavirus,TNF