Immunofluorescence analysis of PAK-1 was done on 70% confluent log phase U87MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with PAK-1 Rabbit Polyclonal Antibody (719300) at 2ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rabbit, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from an internal sequence in the human PAK1 protein. This sequence is identical in human, mouse, and rat PAK1.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||0.5-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PAK proteins are critical effectors that link RhoGTPases to cytoskeleton reorganization and nuclear signaling. PAK proteins, a family of serine/threonine p21-activating kinases, include PAK1, PAK2, PAK3 and PAK4. These proteins serve as targets for the small GTP binding proteins Cdc42 and Rac and have been implicated in a wide range of biological activities. PAK1 regulates cell motility and morphology.
IP-MS enrichment of PAK1 (LFQ intensity): PAK1 was enriched 208-fold from HEK293 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and PAK1 antibody (Part No. 71-9300). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Functional integrity of the T-tubular system in cardiomyocytes depends on p21-activated kinase 1.
71-9300 was used in western blot to identify the role of p21-activated kinase 1 in the T-tubular system in cardiomyocytes
|DeSantiago J,Bare DJ,Ke Y,Sheehan KA,Solaro RJ,Banach K||Journal of molecular and cellular cardiology (60:121)||2013|
PAK1 protein expression in the auditory cortex of schizophrenia subjects.
71-9300 was used in western blot to determine if auditory cortex PAK protein expression in schizophrenia is decreased.
|Deo AJ,Goldszer IM,Li S,DiBitetto JV,Henteleff R,Sampson A,Lewis DA,Penzes P,Sweet RA||PloS one (8:null)||2013|
|Human||Not Cited||Role of group A p21-activated kinases in activation of extracellular-regulated kinase by growth factors.||Beeser A,Jaffer ZM,Hofmann C,Chernoff J||The Journal of biological chemistry (280:36609)||2005|
|Human||Not Cited||p21-activated kinase 1 regulates microtubule dynamics by phosphorylating tubulin cofactor B.||Vadlamudi RK,Barnes CJ,Rayala S,Li F,Balasenthil S,Marcus S,Goodson HV,Sahin AA,Kumar R||Molecular and cellular biology (25:3726)||2005|
|Dynein light chain 1 phosphorylation controls macropinocytosis.||Yang Z,Vadlamudi RK,Kumar R||The Journal of biological chemistry (280:654)||2005|
|Human||Not Cited||Regulation of phosphoglucomutase 1 phosphorylation and activity by a signaling kinase.||Gururaj A,Barnes CJ,Vadlamudi RK,Kumar R||Oncogene (23:8118)||2004|
|Human||Not Cited||Functional inactivation of a transcriptional corepressor by a signaling kinase.||Barnes CJ,Vadlamudi RK,Mishra SK,Jacobson RH,Li F,Kumar R||Nature structural biology (10:622)||2003|
|Filamin is essential in actin cytoskeletal assembly mediated by p21-activated kinase 1.||Vadlamudi RK,Li F,Adam L,Nguyen D,Ohta Y,Stossel TP,Kumar R||Nature cell biology (4:681)||2002|