bs-3314R
[Primary Antibody]
PAK4(Ser474)/PAK5(Ser602)/PAK6(Ser560) Antibody
DATASHEET
Host: Rabbit
Target Protein: PAK4 Ser474/PAK5 Ser602/PAK6 Ser560
Modification Site: Ser560
Clonality: Polyclonal
Isotype: IgG
Entrez Gene: 10298
Source: KLH conjugated synthetic phosphopeptide derived from human PAK4 around the phosphorylation site of PAK4 Ser474 [RK(p-S)LV]
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Store at -20°C for 12 months.
Background:
p21-activated kinases (PAKs) belong to the family of serine/threonine kinases involved in the control of various cellular processes, including the cell cycle, dynamics of the cytoskeleton, apoptosis, oncogenic transformation, and transcription. All PAK family members are characterized by the presence of p21-binding domain. p21-activated kinases are regulated by the small GTP-binding proteins Rac and Cdc42, and lipids, which stimulate autophosphorylation and phosphorylation of exogenous substrates. Serine (Ser-474) is the likely autophosphorylation site in the kinase domain of PAK4 in vivo. Phosphospecific directed against serine 474 detect activated PAK4 on the Golgi membrane when PAK4 is co-expressed with activated Cdc42. Current data strongly implicates PAK-4 in oncogenesis. PAK4 is frequently overexpressed in human tumor cell lines of various tissue origins.
Size: 100ul
Concentration: 1ug/ul
Applications:
WB(1:300-5000)
FCM(1:20-100)
IHC-P(1:200-400)
IF(ICC)(1:50-200)
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
PRODUCT SPECIFIC PUBLICATIONS
- Choi, Sik‐Won, et al. "Repositioning Potential of PAK4 to Osteoclastic Bone Resorption." Journal of Bone and Mineral Research (2015).Read more>>
VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded (Rat skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PAK4(Ser474)/PAK5(Ser602)/PAK6(Ser560)) Polyclonal Antibody, Unconjugated (bs-3314R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PAK4(Ser474)/PAK5(Ser602)/PAK6(Ser560)) Polyclonal Antibody, Unconjugated (bs-3314R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PAK4(Ser474)/PAK5(Ser602)/PAK6(Ser560)) Polyclonal Antibody, Unconjugated (bs-3314R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PAK4(Ser474)/PAK5(Ser602)/PAK6(Ser560)) Polyclonal Antibody, Unconjugated (bs-3314R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse esophageal); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PAK4(Ser474)/PAK5(Ser602)/PAK6(Ser560)) Polyclonal Antibody, Unconjugated (bs-3314R) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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 probed with Phospho-PAK6 (Ser560) Polyclonal Antibody, Unconjugated (bs-3314R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
A-431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Phospho-PAK6 (Ser560)) polyclonal Antibody, Unconjugated (bs-3314R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
A594 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Phospho-PAK6 (Ser560) Antibody(bs-3314R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).