Anti-Phospho-NF-kB p65 (Ser468) Polyclonal Antibody is a Rabbit antibody targeting Phospho-NF-kB p65 (Ser468). Anti-Phospho-NF-kB p65 (Ser468) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. Sample:
LympHnode (Mouse) Lysate at 40 μg
Primary: Anti-Phospho-NFKB p65 (Ser468) (TMAB-01466) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDa
2. Paraformaldehyde-fixed, paraffin embedded (Rat bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-NFKB p65 (Ser468)) Polyclonal Antibody, Unconjugated (TMAB-01466) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-Phospho-NFKB p65 (Ser468) antibody (TMAB-01466), Dilution: 0.2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.
4. Paraformaldehyde-fixed, paraffin embedded (rat lymphoid); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-NFKB p65 (Ser468)) Polyclonal Antibody, Unconjugated (TMAB-01466) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-Phospho-NFKB p65 (Ser468) antibody (TMAB-01466)
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.
6. Blank control: A431. Primary Antibody (green line): Rabbit Anti-Phospho-NFKB p65 (Ser468) antibody (TMAB-01466)
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.
7. Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-Phospho-NFKB p65 (Ser468) antibody (TMAB-01466)
Dilution: 2 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.
8. 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 (Phospho-NFKB p65 (Ser468)) polyclonal Antibody, Unconjugated (TMAB-01466) 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.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:100-200; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-200; FCM: 1μg /test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human NFKBp65 around the phosphorylation site of Ser468

Human

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