Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) Recombinant Rabbit Monoclonal Antibody [SU03-70]
cat.: ET1608-58
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: SU03-70
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 61 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser144 of Human PAK1 aa 121-150 / 545.
Positive control: Hela cell lysate, NIH/3T3 cell lysate, SH-SY5Y cell lysate, Hela, NIH/3T3, human liver carcinoma tissue, human colon carcinoma tissue, mouse brain tissue.
Subcellular location: Cell membrane, ruffle membrane, nucleoplasm, Cytoplasm, focal adhesion, invadopodium, Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: O75914 Human | Q13153 Human | Q13177 Human | O88643 Mouse | Q61036 Mouse | Q8CIN4 Mouse | P35465 Rat | Q62829 Rat | Q64303 Rat
Alternative names: Beta PAK bPAK CDKN1A hPAK3 Mental retardation X linked 30 MRX30 MRX47 Oligophrenin 3 OPHN3 p21 (CDKN1A) activated kinase 3 p21 activated kinase 3 p21 CDKN1A activated kinase 3 P21 protein (Cdc42/Rac) activated kinase 3 PAK-3 PAK3 p21 protein (Cdc42/Rac)-activated kinase 3 PAK3beta Pak65alpha Pak65beta Serine threonine protein kinase PAK 3 Stk4
Images
ET1608-58_1.jpg Fig1: Western blot analysis of Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-58, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: SH-SY5Y cell lysate
ET1608-58_2.jpg Fig2: ICC staining of Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-58, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-58_3.jpg Fig3: ICC staining of Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-58, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1608-58_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-58, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-58_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-58, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-58_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-58, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1608-58_7.jpg Fig7: Flow cytometric analysis of Phospho-PAK1 (S144)+PAK2 (S141)+PAK3 (S139) was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-58, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.