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PAK2 Recombinant Rabbit Monoclonal Antibody [JB41-84]

Usd: 385

Specification

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Catalog# ET7107-79

PAK2 Recombinant Rabbit Monoclonal Antibody [JB41-84]

Application

  • WB

  • IHC-P

  • FC

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

Conjugation

  • unconjugated

Overview

Product Name

PAK2 Recombinant Rabbit Monoclonal Antibody [JB41-84]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N-terminal Human PAK2 .

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IHC-P, FC, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 58 kDa

Positive Control

HeLa cell lysate, HEK-293 cell lysate, MCF7 cell lysate, COS-1 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, A549 cell lysate, human kidney tissue, human skeletal muscle tissue, mouse kidney tissue, mouse skeletal muscle tissue, rat kidney tissue, rat skeletal muscle tissue, A431, SiHa.

Conjugation

unconjugated

Clone Number

JB41-84

RRID

AB_3070787

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200-1:1,000

  • IF-Cell

  • 1:100-1:500

  • IF-Tissue

  • 1:100-1:500

  • FC

  • 1:1,000

Target

Function

Three recently identified isoforms of serine/threonine kinases, designated αPAK p68, βPAK p65 and γPAK p62, have been shown to exhibit a high degree of sequence homology with the S. cerevisiae kinase STE20, involved in pheromone signaling. The α, β, and γPAK isoforms complex specifically with Rac1 and Cdc42 in their active GTP bound state, inhibiting their intrinsic GTPase activity leading to their autophosphorylation. Once phosphorylated and their affinity for Rac/Cdc42 reduced, the PAK isoforms disassociate from the complex to seek downstream substrates. One such putative substrate is MEK kinase, an upstream effector of MEK4 which is involved in the JNK signaling pathway. While the PAK isoforms interact in a GTP-dependent manner with Rac1 and Cdc42, they do not interact with Rho.

Background References

1. Rudel T et al. Membrane and morphological changes in apoptotic cells regulated by caspase-mediated activation of PAK2. Science 276:1571-1574 (1997).

2. Jakobi R et al. Caspase-activated PAK-2 is regulated by subcellular targeting and proteasomal degradation. J Biol Chem 278:38675-38685 (2003).

Sequence Similarity

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.

Tissue Specificity

Ubiquitously expressed. Higher levels seen in skeletal muscle, ovary, thymus and spleen.

Post-translational Modification

Full-length PAK2 is autophosphorylated when activated by CDC42/p21. Following cleavage, both peptides, PAK-2p27 and PAK-2p34, become highly autophosphorylated, with PAK-2p27 being phosphorylated on serine and PAK-2p34 on threonine residues, respectively. Autophosphorylation of PAK-2p27 can occur in the absence of any effectors and is dependent on phosphorylation of Thr-402, because PAK-2p27 is acting as an exogenous substrate.; During apoptosis proteolytically cleaved by caspase-3 or caspase-3-like proteases to yield active PAK-2p34.; Ubiquitinated, leading to its proteasomal degradation.; PAK-2p34 is myristoylated.

Subcellular Location

Cytoplasm, Membrane, Nucleus.

UNIPROT

SwissProt: Q13177 Human

SwissProt: Q8CIN4 Mouse

SwissProt: Q64303 Rat

Synonyms

C-t-PAK2 antibody

CB422 antibody

EC 2.7.11.1 antibody

Gamma PAK antibody

Gamma-PAK antibody

hPAK65 antibody

Kinase antibody

p21 (CDKN1A) activated kinase 2 antibody

p21 (CDKN1A)-activated kinase 2a antibody

p21 activated kinase 2 antibody

Expand

Images

  • Western blot analysis of PAK2 on different lysates with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: COS-1 cell lysate<br />Lane 5: RAW264.7 cell lysate<br />Lane 6: NIH/3T3 cell lysate<br />Lane 7: C6 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 58 kDa<br /><br />Exposure time: 12 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PAK2 on different lysates with Rabbit anti-PAK2 antibody (ET7107-79) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: COS-1 cell lysate
    Lane 5: RAW264.7 cell lysate
    Lane 6: NIH/3T3 cell lysate
    Lane 7: C6 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 58 kDa
    Observed band size: 58 kDa

    Exposure time: 12 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-79) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PAK2 on different lysates with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si PAK2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 58 kDa<br />Observed band size: 58 kDa<br /><br />Exposure time: 30 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of PAK2 on different lysates with Rabbit anti-PAK2 antibody (ET7107-79) at 1/1,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si PAK2 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 58 kDa
    Observed band size: 58 kDa

    Exposure time: 30 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-79) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PAK2 antibody (ET7107-79) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-79) at 1/1,000 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-PAK2 antibody (ET7107-79) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-79) at 1/1,000 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-PAK2 antibody (ET7107-79) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-79) at 1/1,000 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue with Rabbit anti-PAK2 antibody (ET7107-79) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-79) at 1/1,000 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/1,000 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PAK2 antibody (ET7107-79) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-79) at 1/1,000 dilution for 1 hour 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.

  • Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-PAK2 antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-79" style="font-weight: bold;text-decoration: underline;">ET7107-79</a>) at 1/200 dilution for 1 hour 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.

    Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue with Rabbit anti-PAK2 antibody (ET7107-79) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-79) at 1/200 dilution for 1 hour 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.

  • ICC staining PAK2 in A431 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining PAK2 in A431 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • ICC staining PAK2 in SiHa cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining PAK2 in SiHa cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

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