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RabMAb

Anti-Paxillin 抗体 [Y113] (ab32084)

製品の概要

  • 製品名
    Anti-Paxillin antibody [Y113]
    Paxillin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [Y113] to Paxillin
  • 特異性
    ab32084 recognises Paxillin alpha, beta and gamma isoforms.
  • アプリケーション
    適用あり: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Cow, Dog, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Paxillin aa 1-100 (N terminal).

  • ポジティブ・コントロール
    • HeLa cell lysate, human breast carcinoma.
  • 特記事項

     

     

    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 rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32084 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB 1/5000 - 1/10000. Predicted molecular weight: 68 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250.
Flow Cyt Use at an assay dependent concentration.
IP 1/200.

ターゲット情報

  • 機能
    Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion).
  • 配列類似性
    Belongs to the paxillin family.
    Contains 4 LIM zinc-binding domains.
  • 翻訳後修飾
    Phosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens.
  • 細胞内局在
    Cytoplasm > cytoskeleton. Cell junction > focal adhesion.
  • Information by UniProt
  • 参照データベース
  • 別名
    • FLJ16691 antibody
    • FLJ23042 antibody
    • Paired box protein Pax 1 antibody
    • PAX 1 antibody
    • PAX1 antibody
    • PAXI_HUMAN antibody
    • Paxillin alpha antibody
    • Paxillin antibody
    • PXN antibody
    • PXN protein antibody
    see all

画像

  • Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

    Sample: mouse embryonic fibroblasts

    Preparation:

    Fix in 3% PFA in PBS for 30 min at RT

    Primary antibody: Rabbit anti-paxillin Y113 (ab32084), 1:100

    Secondary antibody: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200

    Rhodamine-phalloidin, 1:100

    Nuclei were counterstained with DAPI

     

  • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution ( 10ug/ml) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077) )(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

  • ab32084 staining paxillin in MEF1 cells treated with (S)-(-)-Blebbistatin (ab120491), by ICC/IF. Decreased membrane expression of paxillin correlates with increased concentration of (S)-(-)-Blebbistatin , as described in literature.
    The cells were incubated at 37°C for 2h in media containing different concentrations of ab120491 ( (S)-(-)-Blebbistatin ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32084 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Anti-Paxillin antibody [Y113] (ab32084) at 1/10000 dilution + HeLa cell lysate.

    Predicted band size : 68 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)
  • ab32084 showing positive staining in Normal ovary tissue.

  • Anti-Paxillin antibody [Y113] (ab32084) at 1/5000 dilution + Mouse RAW264.7 whole cell lysate at 20 µg

    Secondary
    An HRP Donkey anti-rabbit IgG polyclonal at 1/10000 dilution

    Predicted band size : 68 kDa

    This image is courtesy of an Abreview submitted by Dr. Alban Gaultier

    Blocking step: 5% Milk for 1 hour at 20°C.\
    Gel: SDS-PAGE

    See Abreview

  • ab32084 showing positive staining in Ovarian carcinoma tissue.

  • ab32084 showing positive staining in Transitional cell carcinoma of kidney tissue.

  • Immunofluorescence analysis of bovine kidney cells, staining Paxillin with ab32084.

    Cells were fixed with paraformaldehyde, permeabilized with 1% Triton X-100 and blocked with 5% BSA for 1 hour. Samples were incubated with primary antibody (1/2500 in 5% BSA) for 1 hour at 25°C. An undiluted AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

    See Abreview

  • ab32084 staining paxillin in MEF1 cells treated with (+/-)-blebbistatin (ab120425), by ICC/IF. Decreased membrane expression of paxillin correlates with increased concentration of (+/-)-blebbistatin, as described in literature.
    The cells were incubated at 37°C for 1h in media containing different concentrations of ab120425 ((+/-)-blebbistatin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32084 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

参考文献

This product has been referenced in:
  • Fantauzzo KA & Soriano P Generation of an immortalized mouse embryonic palatal mesenchyme cell line. PLoS One 12:e0179078 (2017). ICC/IF ; Mouse . Read more (PubMed: 28582446) »
  • Hanga MP  et al. Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system. J Chem Technol Biotechnol 92:1577-1589 (2017). ICC/IF ; Human . Read more (PubMed: 28706339) »

See all 37 Publications for this product

レビューと Q&A

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW264.7)
Specification
RAW264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton + 2% BSA in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C
Username

Dr. Mahesh Shivananjappa

Verified customer

投稿 Apr 16 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HBMEC)
Permeabilization
Yes - 0.1% Triton in PBS
Specification
HBMEC
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Nov 19 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Aug 24 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (HeLa cell)
Specification
HeLa cell
Permeabilization
Yes - 1% Triton X-100 in PBS
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 24 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MDAMB435S)
Specification
MDAMB435S
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Feb 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (rat benign mammary cell line)
Loading amount
20 µg
Specification
rat benign mammary cell line
Gel Running Conditions
Reduced Denaturing (12.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Jan 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Dog Cell lysate - whole cell (MDCK)
Loading amount
10 µg
Specification
MDCK
Gel Running Conditions
Reduced Denaturing (10% tris-glycine gel)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Nov 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HL60)
Loading amount
10 µg
Specification
HL60
Gel Running Conditions
Reduced Denaturing (10% tris-glycine)
Blocking step
Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Nov 02 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Cow Cell (Mardin-Darby bovine kidney cells)
Specification
Mardin-Darby bovine kidney cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% Triton-X-100 (20 min)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

投稿 Oct 16 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Cow Cell lysate - whole cell (MDBK cells)
Loading amount
10 µg
Specification
MDBK cells
Gel Running Conditions
Reduced Denaturing (10 % Tris-Glycine gel)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Oct 16 2012

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

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