Anti-Phospho-PAK6 (Ser560) Polyclonal Antibody is a Rabbit antibody targeting Phospho-PAK6 (Ser560). Anti-Phospho-PAK6 (Ser560) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (rat skin); 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 (Phospho-PAK6 (Ser560)) Polyclonal Antibody, Unconjugated (TMAB-11111) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (Phospho-PAK6 (Ser560)) Polyclonal Antibody, Unconjugated (TMAB-11111) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and 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 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Phospho-PAK6 (Ser560)) Polyclonal Antibody, Unconjugated (TMAB-11111) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (mouse skin); 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 (Phospho-PAK6 (Ser560)) Polyclonal Antibody, Unconjugated (TMAB-11111) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse esophagus); 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 (Phospho-PAK6 (Ser560)) Polyclonal Antibody, Unconjugated (TMAB-11111) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Sample:
Lane 1: Human PANC-1 cell lysates
Lane 2: Human A431 cell lysates
Lane 3: Human A549 cell lysates
Lane 4: Human MCF-7 cell lysates
Primary: Anti-Phospho-PAK6 (Ser560) (TMAB-11111) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 75 kDa
Observed band size: 73 kDa
7. A431 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-PAK6 (Ser560)) polyclonal Antibody, Unconjugated (TMAB-11111) 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 nuclei.
8. Blank control (black line): A549.
Primary Antibody (green line): Rabbit Anti-Phospho-PAK6 (Ser560) antibody (TMAB-11111)
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℃, 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; ICC/IF: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human PAK6 around the phosphorylation site of Ser560

Human

PAK6

Serine/threonine-protein kinase PAK 6

Serine/threonine protein kinase that plays a role in the regulation of gene transcription. The kinase activity is induced by various effectors including AR or MAP2K6/MAPKK6. Phosphorylates the DNA-binding domain of androgen receptor/AR and thereby inhibits AR-mediated transcription. Inhibits also ESR1-mediated transcription. May play a role in cytoskeleton regulation by interacting with IQGAP1. May protect cells from apoptosis through phosphorylation of BAD.