Antibody Type: Rabbit Polyclonal

Application: IHC-P,IHC-Fr,ICC/IF,IF,FCM

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

IgG

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

1. 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-IKK alpha/beta (Ser176 + Ser180)) polyclonal Antibody, Unconjugated (TMAB-01431) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
2. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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-IKK alpha/beta (Ser176 + Ser180)) Polyclonal Antibody, Unconjugated (TMAB-01431) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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-IKK alpha,beta (Ser176 + Ser180)) Polyclonal Antibody, Unconjugated (TMAB-01431) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Blank control (black line): NIH/3T3. Primary Antibody (green line): Rabbit Anti-Phospho-IKK alpha/beta (Ser176 + Ser180) antibody (TMAB-01431)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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.

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

IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human IKK alpha/beta around the phosphorylation site of Ser176/Ser180

Human

1147

NFkB Pathway,Other Kinases,TLR Signaling,IKK,Inflammatory mediators,IKK