Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

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Overview

  • Product name

    Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free
    See all Integrin alpha 2 primary antibodies

  • Description

    Rabbit monoclonal [EPR17338] to Integrin alpha 2 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IP, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    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

  • Positive control

    • 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 tissue, human squamous cell carcinoma of cervix tissue, mouse kidney tissue and rat colon tissues. ICC/IF: Wild-type HAP1, PC-3 and MCF7 cells. Flow Cyt: A549 cells. IP: T-47D whole cell extract.

  • General notes

    ab222377 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.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab222377 in the following tested applications.

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

Application Abreviews Notes
WB Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 129 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
ICC/IF Use at an assay dependent concentration.

This product gave a positive signal in HAP1-ITGA2 wild-type cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min).

Target

  • Function

    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.

  • Sequence similarities

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

  • Domain

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

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 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

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

  • Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

  • Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunocytochemistry/ Immunofluorescence - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

  • Immunoprecipitation - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunoprecipitation - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

  • Flow Cytometry - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Flow Cytometry - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181548).

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

References

This product has been referenced in:

  • Ghosh S  et al. Endoplasmic Reticulum Resident Heat Shock Protein 90 (Hsp90) Isoform Glucose-Regulated Protein 94 (Grp94) Regulates Cell Polarity and Cancer Cell Migration by Affecting Intracellular Transport. J Biol Chem N/A:N/A (2016). WB ; Human . Read more (PubMed: 26872972) »
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