Anti-NF-kB p65 Polyclonal Antibody is a Rabbit antibody targeting NF-kB p65. Anti-NF-kB p65 Polyclonal Antibody can be used in ICC/IF,FCM.

IgG

Human,Mouse (predicted:Zebrafish,Rabbit,Horse,Cow,Pig,Dog,Chicken,Rat)

1. Blank control: mouse splenocytes (blue)
Isotype Control Antibody: Rabbit Igg (orange);
Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue),
Dilution: 1:100 in 1 X PBS containing 0.5% BSA;
Primary Antibody Dilution: 1 μL in 100 μL 1X PBS containing 0.5% BSA (green).
2. Tissue/cell: Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (NFKB p65) polyclonal Antibody, Unconjugated (TMAB-01229) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
3. Blank control: A431. Primary Antibody (green line): Rabbit Anti-NFKB p65 antibody
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
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.
4. Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-NFKB p65 antibody (TMAB-01229)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
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.
5. Tissue/cell: MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (NFKB p65) polyclonal Antibody, Unconjugated (TMAB-01229) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. 4% Paraformaldehyde-fixed Hela (H) cell; Triton X-100 at RT for 20 min; Antibody incubation with (NFKB p65) polyclonal Antibody, unconjugated (TMAB-01229) 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: 1:50-200; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human NFKBp65

Human

RelA (p65),NFkB pathway,Inflammatory mediators,RelA (p65),NFkB pathway,Obesity,Host Immune Response,Host Virus Interaction,Alzheimer's disease,NFkB Pathway