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Knockout Tested Rabbit Recombinant Monoclonal PAK4 antibody. Suitable for IHC-P, WB, IP, Flow Cyt (Intra) and reacts with Human, Recombinant fragment - Human samples.

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Images

Western blot - Anti-PAK4 antibody [EPR28630-82] (AB314765), expandable thumbnail
  • Western blot - Anti-PAK4 antibody [EPR28630-82] (AB314765), expandable thumbnail
  • Western blot - Anti-PAK4 antibody [EPR28630-82] (AB314765), expandable thumbnail
  • Immunoprecipitation - Anti-PAK4 antibody [EPR28630-82] (AB314765), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-PAK4 antibody [EPR28630-82] (AB314765), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBIPFlow Cyt (Intra)ICC/IF
Human
Tested
Tested
Tested
Tested
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Recombinant fragment - Human
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/50
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse, Rat, Recombinant fragment - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Species
Recombinant fragment - Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Recombinant fragment - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/50
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Recombinant fragment - Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat, Recombinant fragment - Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

Serine/threonine protein kinase that plays a role in a variety of different signaling pathways including cytoskeleton regulation, cell migration, growth, proliferation or cell survival. Activation by various effectors including growth factor receptors or active CDC42 and RAC1 results in a conformational change and a subsequent autophosphorylation on several serine and/or threonine residues. Phosphorylates and inactivates the protein phosphatase SSH1, leading to increased inhibitory phosphorylation of the actin binding/depolymerizing factor cofilin. Decreased cofilin activity may lead to stabilization of actin filaments. Phosphorylates LIMK1, a kinase that also inhibits the activity of cofilin. Phosphorylates integrin beta5/ITGB5 and thus regulates cell motility. Phosphorylates ARHGEF2 and activates the downstream target RHOA that plays a role in the regulation of assembly of focal adhesions and actin stress fibers. Stimulates cell survival by phosphorylating the BCL2 antagonist of cell death BAD. Alternatively, inhibits apoptosis by preventing caspase-8 binding to death domain receptors in a kinase independent manner. Plays a role in cell-cycle progression by controlling levels of the cell-cycle regulatory protein CDKN1A and by phosphorylating RAN.

Alternative names

KIAA1142, PAK4, Serine/threonine-protein kinase PAK 4, p21-activated kinase 4, PAK-4

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal PAK4 antibody. Suitable for IHC-P, WB, IP, Flow Cyt (Intra) and reacts with Human, Recombinant fragment - Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR28630-82
Purification technique
Affinity purification Protein A
Specificity

This antibody does not cross-react with human PAK5 and PAK6.

Concentration
Loading...

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.

9 product images

  • Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.
    This target was tested in fresh and frozen LNCaP and Hela, and it was concluded that PAK4 is the easy target for degradation. Lane 3-5 was freshly made and used for western blotting immediately to minimize protein degradation.
    Exposure time: Lane 1, Lane 4-5: 70 seconds; Lane 2: 180 seconds; Lane 3: 26 seconds

    All lanes: Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765) at 1/1000 dilution

    Lanes 1 and 3: LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

    Lanes 2 and 4: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 52 kDa, 68 kDa

  • Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.
    In Western blot, ab314765 was shown to bind specifically to PAK4. Target of interest was observed at 68 kDa and 52 KD in wild-type HEK293T cell lysates (lane 1, wild-type cell line ab255449) with no signal observed at this size in PAK4 knockout cell line (lane 2, knockout cell line Human PAK4 knockout HEK-293T cell line ab266807/ knockout cell lysate Human PAK4 knockout HEK-293T cell lysate ab258560). The band (50 kDa) beneath the target band (68 kDa) is likely to be degraded target fragments. In western blot, anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765) at 1/1000 dilution

    Lane 1: Western blot - Human wild-type HEK-293T cell line (ab255449) at 20 µg

    Lane 2: Western blot - Human PAK4 knockout HEK-293T cell lysate (Human PAK4 knockout HEK-293T cell lysate ab258560) at 20 µg

    Lane 3: PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: Human testis lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 52 kDa, 68 kDa

    Exposure time: 70s

  • Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.
    This antibody does not cross-react with human PAK5 and human PAK6. All the recombinant proteins were made in-house. In western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    All lanes: Western blot - Anti-PAK4 antibody [EPR28630-82] (ab314765) at 1/1000 dilution

    Lane 1: Purified His-tagged human PAK4 recombinant protein at 20 ng

    Lane 2: Purified His-tagged human PAK5 recombinant protein at 100 ng

    Lane 3: Purified His-tagged human PAK6 recombinant protein at 20 ng

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 80 kDa

    Exposure time: 48s

  • Immunoprecipitation - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Immunoprecipitation - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    PAK4 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab314765 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314765 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
    Lane 1: 293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2: ab314765 IP in 293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab314765 in 293T whole cell lysate
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-PAK4 antibody [EPR28630-82] (ab314765) at 1/30 dilution

    All lanes: 293T (human embryonic kidney epithelial cell) whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 15s

  • Flow Cytometry (Intracellular) - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PAK4 KO HEK293T (human PAK4 knockout embryonic kidney epithelial cell, Left) / Parental HEK293T (Right) cells labelling PAK4 with ab314765 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
    Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Immunohistochemical analysis of paraffin-embedded (A) Parental HEK-293T (human embryonic kidney epithelial cell) cell pellet; (B) PAK4 knock-out HEK-293T cell pellet. Tissue labeling PAK4 with ab314765 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Positive staining on (A) parental HEK-293T cell pellet, no staining on (B) PAK4 knock-out HEK-293T cell pellet. The section was incubated with ab314765 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling PAK4 with ab314765 at 1/50 (10.52 ug/ml) dilution, followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Positive staining on human colon carcinoma (PMID: 28542136). The section was incubated with ab314765 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling PAK4 with ab314765 at 1/50 (10.52 ug/ml) dilution, followed by a ready to use rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Positive staining on human breast carcinoma (PMID: 28407679). The section was incubated with ab314765 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK4 antibody [EPR28630-82] (ab314765)

    Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue labeling PAK4 with ab314765 at 1/50 (10.52 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Positive staining on human endometrial carcinoma. The section was incubated with ab314765 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

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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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