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RabMAb

Anti-Integrin alpha 2 抗体 [EPR17338] - C-terminal (ab181548)

製品の概要

  • 製品名

    Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal
    Integrin alpha 2 一次抗体 製品一覧
  • 製品の詳細

    Rabbit monoclonal [EPR17338] to Integrin alpha 2 - C-terminal
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: ICC/IF, IP, Flow Cyt, IHC-P, WBmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Integrin alpha 2 aa 1150 to the C-terminus. The exact sequence is proprietary.
    Database link: P17301

  • ポジティブ・コントロール

    • WB: A549, A431, 293T,T-47D, C6 and NIH/3T3 whole cell lysates, human fetal brain and fetal heart, mouse heart and kidney, and rat spleen tissue lysates. IHC-P: Human colon, human squamous cell carcinoma of cervix, mouse kidney and rat colon tissues. ICC/IF: Wild-type HAP1, PC-3 and MCF7 cells. Flow Cyt: A549 cells. IP: T-47D whole cell extract.
  • 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 ab181548 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
ICC/IF Use a concentration of 1 µg/ml.

This product gave a positive signal in wild-type HAP1 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min).

IP 1/150.
Flow Cyt 1/160.

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

IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 150 kDa (predicted molecular weight: 129 kDa).

ターゲット情報

  • 機能

    Integrin alpha-2/beta-1 is a receptor for laminin, collagen, collagen C-propeptides, fibronectin and E-cadherin. It recognizes the proline-hydroxylated sequence G-F-P-G-E-R in collagen. It is responsible for adhesion of platelets and other cells to collagens, modulation of collagen and collagenase gene expression, force generation and organization of newly synthesized extracellular matrix.
  • 配列類似性

    Belongs to the integrin alpha chain family.
    Contains 7 FG-GAP repeats.
    Contains 1 VWFA domain.
  • ドメイン

    The integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage.
  • 細胞内局在

    Membrane.
  • Information by UniProt
  • 参照データベース

  • 別名

    • BR antibody
    • CD 49b antibody
    • CD49 antigen like family member B antibody
    • CD49 antigen-like family member B antibody
    • CD49b antibody
    • CD49b antigen antibody
    • Collagen receptor antibody
    • DX5 antibody
    • Glycoprotein Ia deficiency included antibody
    • GP Ia antibody
    • GP Ia deficiency, included antibody
    • GPIa antibody
    • HPA 5 included antibody
    • HPA5 included antibody
    • Human platelet alloantigen system 5 antibody
    • Integrin alpha 2 antibody
    • Integrin alpha II antibody
    • Integrin alpha-2 antibody
    • Integrin, alpha 2 (CD49B alpha 2 subunit of VLA 2 receptor) antibody
    • ITA2_HUMAN antibody
    • ITGA2 antibody
    • Platelet alloantigen Br(a), included antibody
    • Platelet antigen Br antibody
    • Platelet glycoprotein GPIa antibody
    • Platelet glycoprotein Ia antibody
    • Platelet glycoprotein Ia/IIa antibody
    • Platelet membrane glycoprotein Ia antibody
    • Platelet receptor for collagen, deficiency of, included antibody
    • Very late activation protein 2 receptor alpha 2 subunit antibody
    • VLA 2 alpha chain antibody
    • VLA 2 antibody
    • VLA 2 subunit alpha antibody
    • VLA-2 subunit alpha antibody
    • VLA2 antibody
    • VLA2 receptor alpha 2 subunit antibody
    • VLAA2 antibody
    see all

画像

  • Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 6 and 10: Integrin alpha 2 knockout HAP1 cell lysate (20 µg)
    Lanes 3, 7 and 11: A431 cell lysate (20 µg)
    Lanes 4, 8 and 12: T47D cell lysate (20 µg)
    Lanes 1, 2, 3 and 4: Green signal from target - ab181548 observed at 150 kDa
    Lanes 5, 6, 7 and 8: Red signal from loading control - ab8245 observed at 37 kDa
    Lanes 9, 10, 11 and 12: Merged (red and green) signal

    ab181548 was shown to specifically react with Integrin alpha 2 when Integrin alpha 2 knockout samples were used. Wild-type and Integrin alpha 2 knockout samples were subjected to SDS-PAGE. ab181548 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane and weak cytoplasmic staining on epithelial cells of human squamous cell carcinoma of cervix tissue is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • ab181548 staining Integrin α2 in wild-type HAP1 cells (top panel) and Integrin α2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab181548 at 1μg/ml concentration and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.


    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • All lanes : Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal (ab181548) at 1/20000 dilution

    Lane 1 : A549 (Human lung carcinoma) whole cell lysates
    Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysates
    Lane 3 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates
    Lane 4 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 129 kDa
    Observed band size: 150 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer 5% NFDM/TBST.

    The increased molecular mass observed is due to glycosylation.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane and weak cytoplasmic staining on epithelial cells of human colon is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling integrin alpha 2 with ab181548 at 1/100 dilution, followed by Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing membrane staining on MCF7 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor®594 secondary antibody) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab181548 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

     

  • All lanes : Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal (ab181548) at 1/5000 dilution

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Mouse kidney tissue lysate
    Lane 3 : Rat spleen tissue lysate
    Lane 4 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 129 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer 5% NFDM/TBST.
    The increased molecular mass observed is due to glycosylation.

  • Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane and weak cytoplasmic staining on epithelial cells of Mouse kidney tubule is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-3 (Human prostate adenocarcinoma cell line) cells labeling integrin alpha 2 with ab181548 at 1/100 dilution, followed by Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing membrane and weakly cytoplasmic staining on PC-3 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor®594 secondary antibody) at 1/500 dilution (red).

    The negative controls are as follows:-
    -ve control 1 - ab181548 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

     

  • Flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling integrin alpha 2 with ab181549 at 1/160 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Integrin alpha 2 with ab181548 at 1/500 dilution followed by Goat Anti-Rabbit HRP (IgG H&L) (ab97051) at 1/500 dilution. Membrane staining on epithelial cells of Rat colon tissue is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal (ab181548) at 1/5000 dilution

    Lane 1 : Human fetal brain whole cell lysates
    Lane 2 : Human fetal heart whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 129 kDa
    Observed band size: 150 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer 5% NFDM/TBST.
    The increased molecular mass observed is due to glycosylation.

  • Integrin alpha 2 was immunoprecipitated from 1mg of T-47D (Human ductal breast epithelial tumor cell line) whole cell extract with ab181548 at 1/150 dilution. Western blot was performed using ab181548 at 1/20,000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: T-47D whole cell extract  Lane 2: PBS instead of T-47D whole cell extract.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

参考文献

This product has been referenced in:

  • Li G  et al. Functional characterization of a potent anti-tumor polysaccharide in a mouse model of gastric cancer. Life Sci 219:11-19 (2019). Read more (PubMed: 30611785) »
  • Wei Q  et al. Low-concentration HCP1 inhibits apoptosis in vascular endothelial cells. Biochem Biophys Res Commun 511:92-98 (2019). Read more (PubMed: 30770100) »
See all 7 Publications for this product

レビューと Q&A

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1-3 of 3 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

投稿 Mar 14 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
colon
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

投稿 Mar 14 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Spleen)
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Formaldehyde

Mr. Simon Cleary

Verified customer

投稿 Jul 02 2015

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