Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Integrin beta 1 antibody [EPR1040Y] (ab134179)

Overview

  • Product name

    Anti-Integrin beta 1 antibody [EPR1040Y]
    See all Integrin beta 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR1040Y] to Integrin beta 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, WB, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Integrin beta 1 aa 50-150. The exact sequence is proprietary.

  • Positive control

    • WB: U937 and HeLa whole cell lysate (ab150035). IHC-P: Human hepatocellular carcinoma and Human brain (endothelial cells) tissues. ICC/IF: Wild-type HAP1 cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab134179 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
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

 HIER is recommended with Tris-EDTA (pH 9.0) or citrate (pH 6.0) buffer.

Flow Cyt 1/100 - 1/1000.

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

WB 1/500. Detects a band of approximately 140-150 kDa (predicted molecular weight: 88 kDa).
ICC/IF 1/250.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Integrins alpha-1/beta-1, alpha-2/beta-1, alpha-10/beta-1 and alpha-11/beta-1 are receptors for collagen. Integrins alpha-1/beta-1 and alpha-2/beta-2 recognize the proline-hydroxylated sequence G-F-P-G-E-R in collagen. Integrins alpha-2/beta-1, alpha-3/beta-1, alpha-4/beta-1, alpha-5/beta-1, alpha-8/beta-1, alpha-10/beta-1, alpha-11/beta-1 and alpha-V/beta-1 are receptors for fibronectin. Alpha-4/beta-1 recognizes one or more domains within the alternatively spliced CS-1 and CS-5 regions of fibronectin. Integrin alpha-5/beta-1 is a receptor for fibrinogen. Integrin alpha-1/beta-1, alpha-2/beta-1, alpha-6/beta-1 and alpha-7/beta-1 are receptors for lamimin. Integrin alpha-4/beta-1 is a receptor for VCAM1. It recognizes the sequence Q-I-D-S in VCAM1. Integrin alpha-9/beta-1 is a receptor for VCAM1, cytotactin and osteopontin. It recognizes the sequence A-E-I-D-G-I-E-L in cytotactin. Integrin alpha-3/beta-1 is a receptor for epiligrin, thrombospondin and CSPG4. Alpha-3/beta-1 may mediate with LGALS3 the stimulation by CSPG4 of endothelial cells migration. Integrin alpha-V/beta-1 is a receptor for vitronectin. Beta-1 integrins recognize the sequence R-G-D in a wide array of ligands. Isoform 2 interferes with isoform 1 resulting in a dominant negative effect on cell adhesion and migration (in vitro). When associated with alpha-7/beta-1 integrin, regulates cell adhesion and laminin matrix deposition. Involved in promoting endothelial cell motility and angiogenesis. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process and the formation of mineralized bone nodules. May be involved in up-regulation of the activity of kinases such as PKC via binding to KRT1. Together with KRT1 and RACK1, serves as a platform for SRC activation or inactivation. Plays a mechanistic adhesive role during telophase, required for the successful completion of cytokinesis. Integrin alpha-3/beta-1 provides a docking site for FAP (seprase) at invadopodia plasma membranes in a collagen-dependent manner and hence may participate in the adhesion, formation of invadopodia and matrix degradation processes, promoting cell invasion. ITGA4:ITGB1 binds to fractalkine (CX3CL1) and may act as its coreceptor in CX3CR1-dependent fractalkine signaling (PubMed:23125415, PubMed:24789099). ITGA4:ITGB1 and ITGA5:ITGB1 bind to PLA2G2A via a site (site 2) which is distinct from the classical ligand-binding site (site 1) and this induces integrin conformational changes and enhanced ligand binding to site 1 (PubMed:18635536, PubMed:25398877). ITGA5:ITGB1 acts as a receptor for fibrillin-1 (FBN1) and mediates R-G-D-dependent cell adhesion to FBN1 (PubMed:12807887, PubMed:17158881).
      Isoform 5: Isoform 5 displaces isoform 1 in striated muscles.
      (Microbial infection) Integrin ITGA2:ITGB1 acts as a receptor for human echoviruses 1 and 8 (PubMed:8411387). Acts as a receptor for cytomegalovirus/HHV-5 (PubMed:20660204). Acts as a receptor for Epstein-Barr virus/HHV-4 (PubMed:17945327). Integrin ITGA5:ITGB1 acts as a receptor for human parvovirus B19 (PubMed:12907437). Integrin ITGA2:ITGB1 acts as a receptor for human rotavirus (PubMed:12941907). Acts as a receptor for mammalian reovirus (PubMed:16501085). In case of HIV-1 infection, integrin ITGA5:ITGB1 binding to extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions (PubMed:10397733).
    • Tissue specificity

      Isoform 1 is widely expressed, other isoforms are generally coexpressed with a more restricted distribution. Isoform 2 is expressed in skin, liver, skeletal muscle, cardiac muscle, placenta, umbilical vein endothelial cells, neuroblastoma cells, lymphoma cells, hepatoma cells and astrocytoma cells. Isoform 3 and isoform 4 are expressed in muscle, kidney, liver, placenta, cervical epithelium, umbilical vein endothelial cells, fibroblast cells, embryonal kidney cells, platelets and several blood cell lines. Isoform 4, rather than isoform 3, is selectively expressed in peripheral T-cells. Isoform 3 is expressed in non-proliferating and differentiated prostate gland epithelial cells and in platelets, on the surface of erythroleukemia cells and in various hematopoietic cell lines. Isoform 5 is expressed specifically in striated muscle (skeletal and cardiac muscle).
    • Sequence similarities

      Belongs to the integrin beta chain family.
      Contains 1 VWFA domain.
    • Post-translational
      modifications

      The cysteine residues are involved in intrachain disulfide bonds.
    • Cellular localization

      Cell membrane, sarcolemma. Cell junction. In cardiac muscle, isoform 5 is found in costameres and intercalated disks and Cell membrane. Cell projection, invadopodium membrane. Cell projection, ruffle membrane. Recycling endosome. Melanosome. Cleavage furrow. Cell projection, lamellipodium. Cell projection, ruffle. Cell junction, focal adhesion. Cell surface. Isoform 2 does not localize to focal adhesions. Highly enriched in stage I melanosomes. Located on plasma membrane of neuroblastoma NMB7 cells. In a lung cancer cell line, in prometaphase and metaphase, localizes diffusely at the membrane and in a few intracellular vesicles. In early telophase, detected mainly on the matrix-facing side of the cells. By mid-telophase, concentrated to the ingressing cleavage furrow, mainly to the basal side of the furrow. In late telophase, concentrated to the extending protrusions formed at the opposite ends of the spreading daughter cells, in vesicles at the base of the lamellipodia formed by the separating daughter cells. Colocalizes with ITGB1BP1 and metastatic suppressor protein NME2 at the edge or peripheral ruffles and lamellipodia during the early stages of cell spreading on fibronectin or collagen. Translocates from peripheral focal adhesions sites to fibrillar adhesions in a ITGB1BP1-dependent manner. Enriched preferentially at invadopodia, cell membrane protrusions that correspond to sites of cell invasion, in a collagen-dependent manner. Localized at plasma and ruffle membranes in a collagen-independent manner.
    • Information by UniProt
    • Database links

    • Alternative names

      • beta1 integrin antibody
      • CD_antigen=CD29 antibody
      • CD29 antibody
      • Fibronectin receptor subunit beta antibody
      • FNRB antibody
      • Glycoprotein IIa antibody
      • GP IIa antibody
      • GPIIA antibody
      • Integrin beta-1 antibody
      • Integrin subunit beta 1 antibody
      • integrin VLA-4 beta subunit antibody
      • Integrin, beta 1 (fibronectin receptor, beta polypeptide, antigen CD29 includes MDF2, MSK12) antibody
      • ITB1_HUMAN antibody
      • ITGB1 antibody
      • MDF2 antibody
      • MSK12 antibody
      • OTTHUMP00000019420 antibody
      • very late activation protein antibody
      • Very late activation protein, beta polypeptide antibody
      • VLA BETA antibody
      • VLA-4 subunit beta antibody
      • VLA-BETA antibody
      • VLAB antibody
      • VLAbeta antibody
      see all

    Images

    • Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 )
      Lanes 2, 6 and 10: Integrin beta 1 knockout HAP1 cell lysate (20 µg)
      Lanes 3, 7 and 11: U87-MG cell lysate (20 µg)
      Lanes 4, 8 and 12: A431 cell lysate (20 µg)
      Lanes 1, 2, 3 and 4: Green signal from target – ab134179 observed at 140kDa
      Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37kDa
      Lanes 9, 10, 11 and 12: Merged (red and green) signal

      ab134179 was shown to specifically react with Integrin beta 1 in wild-type HAP1 cells. No band was observed when Integrin beta 1 knockout samples were examined. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab134179 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1hr at room temperature before imaging.

    • Lane 1: Hap1 wildtype cell lysate (20 µg)

      Lane 2: ITGB1 Hap1 knockout cell lysate (20 µg)

      Lane 3: HeLa wildtype cell lysate (20 µg)

      Lane 4: ITGB1 HeLa knockout cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab134179 observed at 140-150 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab134179 was shown to react with Integrin beta 1 in HeLa wildtype. Loss of signal was observed when knockout sample ab263847 was used. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab134179 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
    • ab134179 staining Integrin beta 1 in wild-type HAP1 cells (top panel) and Integrin beta 1 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), 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 ab134179 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) 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). Nuclear DNA was labelled in blue with DAPI.

    • Overlay histogram showing MCF7 cells stained with ab134179 (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 (ab134179, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a 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.

    • All lanes : Anti-Integrin beta 1 antibody [EPR1040Y] (ab134179) at 1/500 dilution

      Lane 1 : U937 cell lysate
      Lane 2 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 88 kDa

    • Immunohistochemical analysis of paraffin embedded Human hepatocellular carcinoma tissue labelling Integrin beta 1 with ab134179 antibody at a dilution of 1/100. HIER was performed with citrate buffer (pH 6.0).

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

    • Immunohistochemical analysis of paraffin embedded Human brain (endothelial cells) tissue labelling Integrin beta 1 with ab134179 antibody at a dilution of 1/100.

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

    References

    This product has been referenced in:

    • Nassef MZ  et al. Real Microgravity Influences the Cytoskeleton and Focal Adhesions in Human Breast Cancer Cells. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 31261642) »
    • Barnawi R  et al. ß1 Integrin is essential for fascin-mediated breast cancer stem cell function and disease progression. Int J Cancer 145:830-841 (2019). Read more (PubMed: 30719702) »
    See all 6 Publications for this product

    Customer reviews and Q&As

    Answer

    Yes. The antibody is raised against a peptide from the extracellular domain of human integrin beta 1.

    Read More

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