Name: Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y]
Brand: RabMAb
SKU: ab40795
Rating: 4 (4 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP656Y] to PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154)
Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Alkaline Phosphatase APC Carrier Free HRP PE

产品名称

Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154)抗体[EP656Y]
参阅全部 PAK1+PAK2+PAK3 一抗
描述

兔单克隆抗体[EP656Y] to PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154)
宿主

Rabbit
经测试应用

适用于: Flow Cyt (Intra), WB, IHC-P, IP, 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, RAW 264.7 and C6 cell lysates. IHC: Human liver carcinoma, mouse cerebral cortex, rat cerebral cortex. ICC/IF: HeLa cells. IP: HeLa cell lysate. Flow Cyt (intra): NIH/3T3 cell lysate.
常规说明
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
存储溶液

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EP656Y
同种型

IgG
研究领域

The Abpromise guarantee

Abpromise™承诺保证使用ab40795于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
Flow Cyt (Intra)		1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB	(2)	1/10000 - 1/50000. Detects a band of approximately 66 kDa (predicted molecular weight: 65 kDa).
IHC-P	(1)	1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP		1/40.
ICC/IF	(1)	1/250 - 1/500.

说明
Flow Cyt (Intra) 1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/10000 - 1/50000. Detects a band of approximately 66 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/40.
ICC/IF 1/250 - 1/500.

细胞定位

PAK1: Cytoplasm. Cell junction > focal adhesion. Recruited to focal adhesions upon activation. PAK2: Cytoplasm and Nucleus. Cytoplasm > perinuclear region. Membrane. Interaction with ARHGAP10 probably changes PAK-2p34 location to cytoplasmic perinuclear region. Myristoylation changes PAK-2p34 location to the membrane. PAK3: Cytoplasmic
数据库链接
- Entrez Gene: 5058 Human
- Entrez Gene: 5062 Human
- Entrez Gene: 5063 Human
- Entrez Gene: 18479 Mouse
- Entrez Gene: 18481 Mouse
- Entrez Gene: 224105 Mouse
- Entrez Gene: 29431 Rat
- Entrez Gene: 29432 Rat
- Entrez Gene: 29433 Rat
- GenBank: NP_002568.2 Human
- Omim: 300142 Human
- Omim: 602590 Human
- Omim: 605022 Human
- SwissProt: O75914 Human
- SwissProt: Q13153 Human
- SwissProt: Q13177 Human
- SwissProt: O88643 Mouse
- SwissProt: Q61036 Mouse
- SwissProt: Q8CIN4 Mouse
- SwissProt: P35465 Rat
- SwissProt: Q62829 Rat
- SwissProt: Q64303 Rat
- Unigene: 435714 Human
- Unigene: 518530 Human
- Unigene: 234204 Mouse
- Unigene: 260227 Mouse
- Unigene: 3840 Rat
- Unigene: 9149 Rat
see all

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

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
All lanes : 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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
Immunocytochemistry/ Immunofluorescence - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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)
Flow Cytometry (Intracellular) - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in the human cell line NIH/3T3 (mouse embryo) by intracellular 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 Fluorr^® 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)
Immunoprecipitation - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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

Lane 1: HeLa whole cell lysate (10ug)
Lane 2: ab40795 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab40795 in HeLa (human cervix adenocarcinoma) whole cell lysate
Western blot - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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

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

Predicted band size: 65 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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.
Flow Cytometry (Intracellular) - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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.
Western blot - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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

Predicted band size: 65 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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.
Western blot - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (ab40795)

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
All lanes : 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)

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.

Click here to view the general protocols

SDS download

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Datasheet download

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发表研究结果有使用 ab40795？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab40795 被引用在 40 文献中.

Kumar S et al. Low dose radiation upregulates Ras/p38 and NADPH oxidase in mouse colon two months after exposure. Mol Biol Rep 50:2067-2076 (2023). PubMed: 36542238
Ansari MA et al. Impaired Insulin Signaling Alters Mediators of Hippocampal Synaptic Dynamics/Plasticity: A Possible Mechanism of Hyperglycemia-Induced Cognitive Impairment. Cells 12:N/A (2023). PubMed: 37443762
Su N et al. Targeting P21-activated kinase suppresses proliferation and enhances chemosensitivity in T-cell lymphoblastic lymphoma. Blood Sci 5:249-257 (2023). PubMed: 37941919
Kořánová T et al. PAK1 and PAK2 in cell metabolism regulation. J Cell Biochem 123:375-389 (2022). PubMed: 34750857
Voisin A et al. Proteins Associated with Phagocytosis Alteration in Retinal Pigment Epithelial Cells Derived from Age-Related Macular Degeneration Patients. Antioxidants (Basel) 11:N/A (2022). PubMed: 35453399

View all Publications for this product

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1-4 of 4 Abreviews or Q&A

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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 30 2012

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

Blocking step

Serum as blocking agent for 10 minute(s) · Concentration: 10%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 19 2009

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%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 18 2009

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%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Mar 08 2007

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Key features and details

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概述

产品名称

描述

宿主

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Conjugation kits

Isotype control

应用

The Abpromise guarantee

靶标

细胞定位

数据库链接

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (40)

客户评价及客户问答

Western blot abreview for Anti-PAK1 (phospho S144) antibody [EP656Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-PAK1 (phospho S144) antibody [EP656Y]

Western blot abreview for Anti-PAK1 (phospho S144) antibody [EP656Y]

Immunocytochemistry/ Immunofluorescence abreview for Anti-PAK1 (phospho S144) antibody [EP656Y]