Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) 抗体 [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
Related conjugates and formulations
製品の概要
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製品名
Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y]
PAK1+PAK2+PAK3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP656Y] to PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human PAK1 (phospho S144). The exact sequence is proprietary.
Database link: Q13153 -
ポジティブ・コントロール
- 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.
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特記事項
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP656Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Alexa Fluor® 647 Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] (ab203640)
- Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (ab239830)
- Alexa Fluor® 568 Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP65 (ab312483)
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Conjugation kits
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab40795の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
1/120.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (2) |
1/10000 - 1/50000. Detects a band of approximately 66 kDa (predicted molecular weight: 65 kDa).
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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.
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IP |
1/40.
|
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ICC/IF | (1) |
1/250 - 1/500.
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特記事項 |
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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. |
ターゲット情報
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細胞内局在
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 -
参照データベース
- 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
see all
画像
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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 minuteBlocking and dilution buffer: 5% NFDM/TBST.
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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
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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 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)
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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 -
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
All lanes : 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.
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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.
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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 -
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.
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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
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (35)
ab40795 は 35 報の論文で使用されています。
- 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
- Kuželová K et al. Group I p21-activated kinases in leukemia cell adhesion to fibronectin. Cell Adh Migr 15:18-36 (2021). PubMed: 33464167
- Davidson A et al. A kinase-independent function of PAK is crucial for pathogen-mediated actin remodelling. PLoS Pathog 17:e1009902 (2021). PubMed: 34460869
- Lee SH et al. Haploinsufficiency of Cyfip2 Causes Lithium-Responsive Prefrontal Dysfunction. Ann Neurol 88:526-543 (2020). PubMed: 32562430