Recombinant Anti-PAK1 antibody [EPR20048] (ab223849)
![Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-309606-anti-pak1-antibody-epr20048-western-blot.jpg "Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20048] to PAK1
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PAK1 antibody [EPR20048]
See all PAK1 primary antibodies -
Description
Rabbit monoclonal [EPR20048] to PAK1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: His-tagged human PAK1 (aa1-250) recombinant protein; HeLa, SK-OV-3, SH-SY5Y, HEK-293T, NIH/3T3 and PC-12 whole cell lysates; Human fetal brain lysate; Mouse and rat brain lysates. ICC/IF: SH-SY5Y and HeLa cells. Flow Cyt (intra): SH-SY5Y and HeLa cells. IP: SH-SY5Y whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.1% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20048 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab223849 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 61 kDa).
In WB this antibody showed weak staining of PAK1 on HeLa cell lysate |
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| ICC/IF |
1/100.
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| IP |
1/30.
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| Flow Cyt (Intra) |
1/50.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 61 kDa). In WB this antibody showed weak staining of PAK1 on HeLa cell lysate |
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ICC/IF
1/100. |
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IP
1/30. |
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Flow Cyt (Intra)
1/50. |
Target
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Function
The activated kinase acts on a variety of targets. Likely to be the GTPase effector that links the Rho-related GTPases to the JNK MAP kinase pathway. Activated by CDC42 and RAC1. Involved in dissolution of stress fibers and reorganization of focal complexes. Involved in regulation of microtubule biogenesis through phosphorylation of TBCB. Activity is inhibited in cells undergoing apoptosis, potentially due to binding of CDC2L1 and CDC2L2. -
Sequence similarities
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.
Contains 1 CRIB domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated when activated by CDC42/p21 and RAC1. -
Cellular localization
Cytoplasm. Cell junction > focal adhesion. Recruited to focal adhesions upon activation. - Information by UniProt
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Database links
- Entrez Gene: 5058 Human
- Entrez Gene: 18479 Mouse
- Entrez Gene: 29431 Rat
- Omim: 602590 Human
- SwissProt: Q13153 Human
- SwissProt: O88643 Mouse
- SwissProt: P35465 Rat
- Unigene: 435714 Human
see all -
Alternative names
- Alpha PAK antibody
- Alpha-PAK antibody
- MGC130000 antibody
see all
Images
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Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)All lanes : Anti-PAK1 antibody [EPR20048] (ab223849) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PAK1 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 61 kDaLanes 1 - 2: Merged signal (red and green). Green - ab223849 observed at 61 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab223849 was shown to specifically react with PAK1 in wild-type HAP1 cells as signal was lost in PAK1 knockout cells. Wild-type and PAK1 knockout samples were subjected to SDS-PAGE. Ab223849 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286958-anti-pak1-antibody-epr20048-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PAK1 antibody [EPR20048] (ab223849)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labeling PAK1 with ab223849 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SH-SY5Y cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286956-anti-pak1-antibody-epr20048-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-PAK1 antibody [EPR20048] (ab223849)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cell line labeling PAK1 with ab223849 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary
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![Immunoprecipitation - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286954-anti-pak1-antibody-epr20048-immunoprecipitation.jpg)
Immunoprecipitation - Anti-PAK1 antibody [EPR20048] (ab223849)PAK1 was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate with ab223849 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab223849 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: SH-SY5Y whole cell lysate 10 μg (Input).
Lane 2: ab223849 IP in SH-SY5Y whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223849 in SH-SY5Y whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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![Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-397478-anti-pak1-antibody-epr20048-western-blot-hela-edited-lysates.jpg)
Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)All lanes : Anti-PAK1 antibody [EPR20048] (ab223849) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PAK1 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab223849 observed at 65 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab223849 was shown to react with PAK1 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 editedcell line ab264889 (CRISPR/Cas9 editedcell lysate ab257572) lane below 65kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PAK1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab223849 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286960-anti-pak1-antibody-epr20048-western-blot.jpg)
Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)All lanes : Anti-PAK1 antibody [EPR20048] (ab223849) at 1/1000 dilution
Lane 1 : SK-OV-3 (human ovarian cancer epithelial cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : Human fetal brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 3 minutes; Lane 2: 30 seconds; Lane 3: 3 seconds; Lane 4: 15 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286959-anti-pak1-antibody-epr20048-western-blot.jpg)
Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)All lanes : Anti-PAK1 antibody [EPR20048] (ab223849) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : SH-SY5Y (human euroblastoma cell line from bone marrow) whole cell lysate
Lane 4 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 5 seconds; Lane 2: 3 seconds; Lane 3: 3 minutes; Lane 4: 1 minutes.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286961-anti-pak1-antibody-epr20048-western-blot.jpg)
Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)All lanes : Anti-PAK1 antibody [EPR20048] (ab223849) at 1/1000 dilution
Lane 1 : His-tagged human PAK1 (aa1-250) recombinant protein
Lane 2 : His-tagged human PAK2 (aa1-250) recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 61 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 1 minute; Lane 2: 3 minutes.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286957-anti-pak1-antibody-epr20048-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PAK1 antibody [EPR20048] (ab223849)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling PAK1 with ab223849 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-286955-anti-pak1-antibody-epr20048-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-PAK1 antibody [EPR20048] (ab223849)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling PAK1 with ab223849 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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![Anti-PAK1 antibody [EPR20048] (ab223849)](https://www.abcam.com/ps/products/223/ab223849/Images/ab223849-11-benefits-of-recombinant-antibodies.png)
Anti-PAK1 antibody [EPR20048] (ab223849)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (7)
ab223849 has been referenced in 7 publications.
- Kuželová K et al. Group I p21-activated kinases in leukemia cell adhesion to fibronectin. Cell Adh Migr 15:18-36 (2021). PubMed: 33464167
- Li H et al. Exosomes containing miR-451a is involved in the protective effect of cerebral ischemic preconditioning against cerebral ischemia and reperfusion injury. CNS Neurosci Ther 27:564-576 (2021). PubMed: 33533575
- Chen YH et al. Mesenchymal Stem Cell-Derived Exosomes Induced by IL-1ß Attenuate Urethral Stricture Through Let-7c/PAK1/NF-?B-Regulated Macrophage M2 Polarization. J Inflamm Res 14:3217-3229 (2021). PubMed: 34285545
- Lin J et al. MiR-23a induced the activation of CDC42/PAK1 pathway and cell cycle arrest in human cov434 cells by targeting FGD4. J Ovarian Res 13:90 (2020). PubMed: 32772928
- Yao D et al. Design, synthesis and biological evaluation of novel tetrahydrothieno [2,3-c]pyridine substitued benzoyl thiourea derivatives as PAK1 inhibitors in triple negative breast cancer. J Enzyme Inhib Med Chem 35:1524-1538 (2020). PubMed: 32752894
- Grebenová D et al. PAK1, PAK1?15, and PAK2: similarities, differences and mutual interactions. Sci Rep 9:17171 (2019). PubMed: 31748572
- Liu C et al. Epigenetically upregulated GEFT-derived invasion and metastasis of rhabdomyosarcoma via epithelial mesenchymal transition promoted by the Rac1/Cdc42-PAK signalling pathway. EBioMedicine 50:122-134 (2019). PubMed: 31761617