Anti-Phospho-Cytokeratin 8 (Ser431) Polyclonal Antibody

Anti-Phospho-Cytokeratin 8 (Ser431) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Cytokeratin 8 (Ser431). Anti-Phospho-Cytokeratin 8 (Ser431) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse (predicted:Rat,Horse)

1. Sample: Stomach (Mouse) Lysate at 40 μg
Primary: Anti-phospho-Cytokeratin 8 (Ser431) (TMAB-10569) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 53 kD
2. Paraformaldehyde-fixed, paraffin embedded (Human colon cancer); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (p-Cytokeratin 8 (Ser431)) Polyclonal Antibody, Unconjugated (TMAB-10569) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
3. Blank control: MCF-7.
Primary Antibody (green line): Rabbit Anti-phospho-Cytokeratin 8 (Ser431) antibody (TMAB-10569)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 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℃. 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. Acquisition of 20,000 events was performed.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human CK8 around the phosphorylation site of Ser431

Human

KRT8

Keratin, type II cytoskeletal 8

Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.