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RabMAb

Anti-Integrin alpha 5 抗体 [EPR7854] - Low endotoxin, Azide free (ab221606)

製品の概要

  • 製品名

    Anti-Integrin alpha 5 antibody [EPR7854] - Low endotoxin, Azide free
    Integrin alpha 5 一次抗体 製品一覧
  • 製品の詳細

    Rabbit monoclonal [EPR7854] to Integrin alpha 5 - Low endotoxin, Azide free
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, IHC-P, ICC/IF, IP, Flow Cytmore 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) (C terminal)

  • ポジティブ・コントロール

    • IHC-P: Human kidney, human cerebral cortex, mouse cerebral cortex, and rat cerebral cortex tissues. ICC/IF: U937, MCF7 and wild-type HAP1 cells. Flow Cyt: HeLa cells. IP: HeLa cells.
  • 特記事項

    ab221606 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

    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.

    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 ab221606 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 Use at an assay dependent concentration. Detects a band of approximately 19, 150 kDa (predicted molecular weight: 115 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能

    Integrin alpha-5/beta-1 is a receptor for fibronectin and fibrinogen. It recognizes the sequence R-G-D in its ligands. In case of HIV-1 infection, the interaction with extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions.
  • 配列類似性

    Belongs to the integrin alpha chain family.
    Contains 7 FG-GAP repeats.
  • 細胞内局在

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

  • 別名

    • CD49 antigen-like family member E antibody
    • CD49e antibody
    • Fibronectin receptor subunit alpha antibody
    • Fibronectin receptor, alpha subunit antibody
    • FNRA antibody
    • Integrin alpha 5 (fibronectin receptor alpha) antibody
    • Integrin alpha-5 antibody
    • Integrin alpha-5 light chain antibody
    • Integrin alpha-F antibody
    • Integrin, alpha 5 (fibronectin receptor, alpha polypeptide) antibody
    • ITA5_HUMAN antibody
    • Itga5 antibody
    • Very late activation protein 5, alpha subunit antibody
    • VLA-5 antibody
    • VLA5 antibody
    • VLA5A antibody
    see all

画像

  • ab150361 staining Integrin alpha 5 in MCF7 cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Tween 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 ab150361 at 1μg/ml concentration and ab7291 at 1μg/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 2 μg/ml (showed in pseudocolour red) . Nuclear DNA was labelled in blue with DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

  • ab150361 (Purified) at 1:20 dilution (1 µg) immunoprecipitating Integrin alpha 5 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
    Lane 2 (+): ab150361 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab150361 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1:5000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361)
  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Integrin alpha 5 with purified ab150361 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361)
  • Immunocytochemistry/ Immunofluorescence analysis of U937 (Human histiocytic lymphoma monocyte) cells labeling Integrin alpha 5 with purified ab150361 at 1:50 dilution (4.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue sections labeling Integrin alpha 5 with purified ab150361 at 1:400 dilution (0.54 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue sections labeling Integrin alpha 5 with purified ab150361 at 1:400 dilution (0.54 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue sections labeling Integrin alpha 5 with purified ab150361 at 1:400 dilution (0.54 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361)
  • ab150361 staining Integrin alpha 5 in wild-type HAP1 cells (top panel) and ITGA5 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Tween 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 ab150361 at 1/250 dilution and ab7291 at 1μg/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 2 μg/ml (showed in pseudocolour red) . Nuclear DNA was labelled in blue with DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U937 (Human histiocytic lymphoma) cells labeling Integrin alpha 5 with unpurified ab150361 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing mainly membrane staining on U937 cell line. The nuclear counterstain is DAPI (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

  • Overlay histogram showing HeLa cells stained with unpurified ab150361 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab150361, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor 488 IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

  • Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Integrin alpha 5 with unpurified ab150361 antibody at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin embedded Human Vessels tissue using unpurified ab150361 showing +ve staining.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin embedded Human Brain vessels tissue using unpurified ab150361 showing +ve staining.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin embedded Human Skeletal muscle vessel tissue using unpurified ab150361 showing +ve staining.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin embedded Normal Human Tonsil tissue using unpurified ab150361 showing +ve staining.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin embedded Normal Human Uterus tissue using unpurified ab150361 showing +ve staining.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150361).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

参考文献

This product has been referenced in:

  • McMillan KJ  et al. Atypical parkinsonism-associated retromer mutant alters endosomal sorting of specific cargo proteins. J Cell Biol 214:389-99 (2016). Read more (PubMed: 27528657) »
See 1 Publication for this product

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