Antibody Type: Recombinant Rabbit Monoclonal

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

Reactivity: Human (predicted:Mouse,Rat,Pig,Cow,Rabbit,Sheep)

IgG

7N349

Human (predicted:Mouse,Rat,Pig,Cow,Rabbit,Sheep)

1. Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-phospho-IKB alpha (Ser32) antibody (TMAB-01424)
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. Sample:
Lane 1: SK-Br-3 cell lysates
Primary: Anti-phospho-IKB alpha (Ser32) (TMAB-01424) at 1/500 dilution
Secondary: Goat Anti-Rabbit IgG-HRP at 1:5000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
3. 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-IKB alpha (S32)) monoclonal Antibody, Unconjugated (TMAB-01424) 1:50, 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.

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

WB: 1:500-1000; IHC-P: 1:100-500; IHC-Fr: 1:50-200; ICC/IF: 1:50; IF: 1:50-200; FCM: 2ug/Test

Monoclonal

KLH conjugated Synthesised phosphopeptide: human IKB alpha around the phosphorylation site of Ser32

Human

4792

NFkB pathway,NFkB pathway,TLR Signaling,NFkB Pathway