Invitrogen

PAK1 Polyclonal Antibody

Advanced Verification

IP-mass spectrometry View Details

6 Published Figures

9 References

View all (27) PAK1 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

PAK1 Antibody in Immunofluorescence (IF) — 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 (Product # 71-9300) at 2 µg/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 (Product # 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.

PAK1 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of Mouse Skeletal Muscle (Lane 1), MDA-MB-231 (lane 2), MCF7 (lane 3), CaCo-2 (lane 4) and Jurkat (lane 5). The blots were probed with Anti-PAK1 Rabbit Polyclonal Antibody (Product # 71-9300, 0.5-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Two bands ~ 61 and 68 kDa corresponding to PAK1 were observed across cell lines and tissues tested expect in MDA-MB-231. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

PAK1 Antibody in Knockdown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. Caco-2 cells were transfected with PAK1 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-PAK1 Polyclonal Antibody (Product # 71-9300). {KD}

PAK1 Antibody in IP-MS — 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 (Product # 90409) and PAK1 antibody (Product # 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}

PAK1 Antibody — Figure 3 Effects of postmortem interval on PAK1 expression. A ) Example western blot of PAK1 and Tubulin in mice in which the interval from sacrifice to brain harvesting was experimentally varied between 0 and 48 hours. MW, Molecular Weight Markers. B ) Optical densities for PAK1 and Tubulin in mice in which the interval from sacrifice to brain harvesting was experimentally varied between 0 and 48 hours. Two sets of mice (each set containing one mouse for each PMI) were tested. Sets were run concurrently on separate gels, and the run was repeated. Mean (SEM) values for the four observations at each time point are shown. Time points sharing the same superscript letter differ significantly.

Product Details

Applications

Tested Dilution

Publications

ELISA (ELISA)

Assay Dependent

Immunocytochemistry (ICC)

1-2 µg/mL

Immunofluorescence (IF)

1-2 µg/mL

View 5 publications 5 publications

Immunoprecipitation (IP)

Assay Dependent

Western Blot (WB)

0.5-2 µg/mL

View 4 publications 4 publications

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human, Mouse, Rabbit

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide derived from an internal sequence in the human PAK1 protein. This sequence is identical in human, mouse, and rat PAK1.

Conjugate

Unconjugated

Form

Liquid

Concentration

0.25 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7.4

Contains

0.1% sodium azide

Storage conditions

-20°C

RRID

AB_2534005

Target Information

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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: activated protein kinase alpha; ADRB2; alpha pak; Alpha-PAK; CDC42/RAC effector kinase PAK-A; EC 2.7.11.1; kinase PAK1; MGC130000; MGC130001; p21 (CDKN1A)-activated kinase 1; p21 protein (Cdc42/Rac)-activated kinase 1; p21-activated kinase 1; p21/Cdc42/Rac1-activated kinase 1 (STE20 homolog, yeast); p21/Cdc42/Rac1-activated kinase 1 (yeast Ste20-related); p65-PAK; p68-PAK; PAK 1; PAK-1; PAK1B; Protein kinase MUK2; Serine/threonine-protein kinase PAK 1; STE20 homolog, yeast

Gene Aliases: AW045634; PAK-1; PAK1; Paka; PAKalpha

UniProt ID: (Human) Q32M54, (Rat) P35465

Entrez Gene ID: (Human) 5058, (Mouse) 18479, (Rat) 29431

Function(s)