重组
RabMAb

Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154)抗体[EP656Y] (ab40795)

概述

  • 产品名称
    Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154)抗体[EP656Y]
  • 描述
    兔单克隆抗体[EP656Y] to PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154)
  • 特异性
    ab203958 recognises p21-activated kinase 1 (PAK1).
  • 经测试应用
    适用于: WB, IHC-P, IP, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human PAK1 (phospho S144). The exact sequence is proprietary.
    Database link: Q13153

  • 阳性对照
    • WB: MCF7, HeLa, RAW264.7 and C6 cell lysates. IHC: Human liver carcinoma, mouse cerebral cortex, rat cerebral cortex. ICC/IF: HeLa cells. IP: HeLa cell lysate. Flow Cyt: NIH/3T3 cell lysate.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

     

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

性能

应用

Our Abpromise guarantee covers the use of ab40795 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/10000 - 1/50000. Detects a band of approximately 66 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/100 - 1/500.
IP 1/40.
Flow Cyt 1/120.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/250 - 1/500.

靶标

Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] 图像

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795) at 1/1000 dilution

    Lane 1 : MCF7, grown in serum-free media overnight, whole cell lysate
    Lane 2 : MCF7, grown in serum-free media overnight, then treated with EGF 1µg/ml for 10min, whole cell lysate
    Lane 3 : MCF7, grown in serum-free media overnight, then treated with EGF 1µg/ml for 10min, whole cell lysate. The membrane was incubated with phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 65 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    Blocking and dilution buffer: 5% NFDM/TBST.

  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in rat cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary

  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in HeLa (human cervix adenocarcinoma) cells, treated and untreated with Lambda Protein Phosphtase 31℃ for 5h by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab75748 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

     

  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in the human cell line NIH/3T3 (mouse embryo) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/120. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • ab40795 immunoprecipitating PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154). 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
    Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab40795 in HeLa (human cervix adenocarcinoma) whole cell lysate

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795) at 1/2000 dilution

    Lane 1 : HeLa cell lysate with None
    Lane 2 : HeLa cell lysate with PAK2 (pS141)
    Lane 3 : HeLa cell lysate with PAK2 non-phospho
    Lane 4 : HeLa cell lysate with PAK3 (pS154)
    Lane 5 : HeLa cell lysate with PAK3 non-phospho


    Predicted band size : 65 kDa
  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795) at 1/50000 dilution

    Lane 1 : C6 (rat glioma) whole cell lysate - treated with phosphatase
    Lane 2 : C6 (rat glioma) whole cell lysate - untreated

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 65 kDa
  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in mouse cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Overlay histogram showing HeLa cells stained with unpurified ab40795 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40795, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795) at 1/10000 dilution

    Lane 1 : HeLa whole cell lysate - untreated
    Lane 2 : HeLa whole cell lysate - treated with phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 65 kDa
  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in human liver carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

     

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795) at 1/10000 dilution

    Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate - treated with phosphatase
    Lane 2 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate - untreated

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 65 kDa

Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)参考文献

This product has been referenced in:
  • Lin XJ  et al. hsa-miR-485-5p reverses epithelial to mesenchymal transition and promotes cisplatin-induced cell death by targeting PAK1 in oral tongue squamous cell carcinoma. Int J Mol Med 40:83-89 (2017). WB ; Human . Read more (PubMed: 28535002) »
  • Schelle I  et al. Role of p38alpha/beta MAP Kinase in Cell Susceptibility to Clostridium sordellii Lethal Toxin and Clostridium difficile Toxin B. Toxins (Basel) 9:N/A (2016). WB . Read more (PubMed: 28025502) »

See all 15 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116)
Loading amount
40 µg
Specification
HCT116
Gel Running Conditions
Non-reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (normal tissue array (including breast/colon/spleen)
Specification
normal tissue array (including breast/colon/spleen
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 10%
Username

Abcam user community

Verified customer

提交于 May 19 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast)
Loading amount
20 µg
Specification
Breast
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Username

Abcam user community

Verified customer

提交于 May 18 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (cultured hippocampal astrocytes)
Specification
cultured hippocampal astrocytes
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1%
Username

Abcam user community

Verified customer

提交于 Mar 08 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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