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Rabbit Recombinant Monoclonal PAK1 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 11 publications.

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Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.1% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Rat
Expected
Tested
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

In WB this antibody showed weak staining of PAK1 on HeLa cell lysate

Species

Rat

Dilution info

1/1000

Notes

In WB this antibody showed weak staining of PAK1 on HeLa cell lysate

Species

Human

Dilution info

1/1000

Notes

In WB this antibody showed weak staining of PAK1 on HeLa cell lysate

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Protein kinase involved in intracellular signaling pathways downstream of integrins and receptor-type kinases that plays an important role in cytoskeleton dynamics, in cell adhesion, migration, proliferation, apoptosis, mitosis, and in vesicle-mediated transport processes (PubMed:10551809, PubMed:11896197, PubMed:12876277, PubMed:14585966, PubMed:15611088, PubMed:17726028, PubMed:17989089, PubMed:30290153). Can directly phosphorylate BAD and protects cells against apoptosis (By similarity). Activated by interaction with CDC42 and RAC1 (PubMed:8805275, PubMed:9528787). Functions as GTPase effector that links the Rho-related GTPases CDC42 and RAC1 to the JNK MAP kinase pathway (PubMed:8805275, PubMed:9528787). Phosphorylates and activates MAP2K1, and thereby mediates activation of downstream MAP kinases (By similarity). Involved in the reorganization of the actin cytoskeleton, actin stress fibers and of focal adhesion complexes (PubMed:9395435, PubMed:9032240). Phosphorylates the tubulin chaperone TBCB and thereby plays a role in the regulation of microtubule biogenesis and organization of the tubulin cytoskeleton (PubMed:15831477). Plays a role in the regulation of insulin secretion in response to elevated glucose levels (PubMed:22669945). Part of a ternary complex that contains PAK1, DVL1 and MUSK that is important for MUSK-dependent regulation of AChR clustering during the formation of the neuromuscular junction (NMJ) (By similarity). Activity is inhibited in cells undergoing apoptosis, potentially due to binding of CDC2L1 and CDC2L2 (PubMed:12624090). Phosphorylates MYL9/MLC2 (By similarity). Phosphorylates RAF1 at 'Ser-338' and 'Ser-339' resulting in: activation of RAF1, stimulation of RAF1 translocation to mitochondria, phosphorylation of BAD by RAF1, and RAF1 binding to BCL2 (PubMed:11733498). Phosphorylates SNAI1 at 'Ser-246' promoting its transcriptional repressor activity by increasing its accumulation in the nucleus (PubMed:15833848). In podocytes, promotes NR3C2 nuclear localization (By similarity). Required for atypical chemokine receptor ACKR2-induced phosphorylation of LIMK1 and cofilin (CFL1) and for the up-regulation of ACKR2 from endosomal compartment to cell membrane, increasing its efficiency in chemokine uptake and degradation (PubMed:23633677). In synapses, seems to mediate the regulation of F-actin cluster formation performed by SHANK3, maybe through CFL1 phosphorylation and inactivation (By similarity). Plays a role in RUFY3-mediated facilitating gastric cancer cells migration and invasion (PubMed:25766321). In response to DNA damage, phosphorylates MORC2 which activates its ATPase activity and facilitates chromatin remodeling (PubMed:23260667). In neurons, plays a crucial role in regulating GABA(A) receptor synaptic stability and hence GABAergic inhibitory synaptic transmission through its role in F-actin stabilization (By similarity). In hippocampal neurons, necessary for the formation of dendritic spines and excitatory synapses; this function is dependent on kinase activity and may be exerted by the regulation of actomyosin contractility through the phosphorylation of myosin II regulatory light chain (MLC) (By similarity). Along with GIT1, positively regulates microtubule nucleation during interphase (PubMed:27012601). Phosphorylates FXR1, promoting its localization to stress granules and activity (PubMed:20417602).

Alternative names

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Rabbit Recombinant Monoclonal PAK1 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 11 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR20048

Purification technique

Affinity purification Protein A

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • Western blot - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)

    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 knockout cell line ab264889 (knockout cell lysate ab257572) lane below 65kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and PAK1 knockout 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.

    All lanes: Western blot - Anti-PAK1 antibody [EPR20048] (AB223849) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: PAK1 knockout HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 61 kDa

    Observed band size: 65 kDa

  • Western blot - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)

    Lanes 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.

    All lanes: Western blot - Anti-PAK1 antibody [EPR20048] (AB223849) at 1/1000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: PAK1 knockout HAP1 whole cell lysate at 20 µg

    Predicted band size: 61 kDa

  • Immunoprecipitation - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    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.

    All lanes: Immunoprecipitation - Anti-PAK1 antibody [EPR20048] (AB223849)

    Predicted band size: 61 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    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.

  • Western blot - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)

    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.

    All lanes: Western blot - Anti-PAK1 antibody [EPR20048] (AB223849) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: PAK1 CRISPR/Cas9 edited HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 61 kDa

    Observed band size: 65 kDa

  • Western blot - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)

    Exposure time : Lane 1: 3 minutes; Lane 2: 30 seconds; Lane 3: 3 seconds; Lane 4: 15 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-PAK1 antibody [EPR20048] (AB223849) at 1/1000 dilution

    Lane 1: SK-OV-3 (human ovarian cancer epithelial cell line) whole cell lysate at 20 µg

    Lane 2: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg

    Lane 3: PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

    Lane 4: Human fetal brain lysate at 20 µg

    Secondary

    Lanes 1 - 3: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution

    Lane 4: Western blot - 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

  • Western blot - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)

    Exposure time : Lane 1: 5 seconds; Lane 2: 3 seconds; Lane 3: 3 minutes; Lane 4: 1 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-PAK1 antibody [EPR20048] (AB223849) at 1/1000 dilution

    Lane 1: Mouse brain lysate at 10 µg

    Lane 2: Rat brain lysate at 10 µg

    Lane 3: SH-SY5Y (human euroblastoma cell line from bone marrow) whole cell lysate at 10 µg

    Lane 4: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - 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

  • Western blot - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    Western blot - Anti-PAK1 antibody [EPR20048] (ab223849)

    Exposure time : Lane 1: 1 minute; Lane 2: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-PAK1 antibody [EPR20048] (AB223849) at 1/1000 dilution

    Lane 1: His-tagged human PAK1 (aa1-250) recombinant protein at 0.01 µg

    Lane 2: His-tagged human PAK2 (aa1-250) recombinant protein at 0.01 µg

    Secondary

    All lanes: Western blot - 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

  • Immunocytochemistry/ Immunofluorescence - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    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.

  • Flow Cytometry (Intracellular) - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    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.

  • Flow Cytometry (Intracellular) - Anti-PAK1 antibody [EPR20048] (ab223849), expandable thumbnail

    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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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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