Validated using a knockout cell line

Anti-Integrin beta 1 antibody [12G10] (ab30394)

Reviews (7)Q&A (20)References (51)

Overview

  • Product name

    Anti-Integrin beta 1 antibody [12G10]
    See all Integrin beta 1 primary antibodies

  • Description

    Mouse monoclonal [12G10] to Integrin beta 1

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, ELISA, WBmore details

  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat

  • Immunogen

    Full length native protein (purified) corresponding to Integrin beta 1. Purified from HT1080 fibrosarcoma cell extract.

  • Epitope

    The 12G10 epitope has been identified to be within the von Willebrand factor type A domain of the ß-subunit (PubMed ID: 15632175).

  • Positive control

    • WB: HT1080 whole cell lysate. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • General notes

    This monoclonal antibody to integrin beta 1 has been knockout validated in ICC/IF and flow cytometry. The expected signal was observed in wild type cells and was not seen in knockout cells.

    The 12G10 monoclonal has been shown to increase ligand binding and enhances α5β1 integrin-fibronectin interactions (PubMed ID: 7537221). The antibody promotes cell attachment and spreading when binding to α5β1 integrin but inhibits these processes when binding to α4β1 integrin (PubMed ID: 15632175). 

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or  conjugation for your experiments, please contact orders@abcam.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab30394 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
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ELISA Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 88 kDa).

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

    • Western blot - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Western blot - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Anti-Integrin beta 1 antibody [12G10] (ab30394) at 5 µg/ml + HT 1080 (Human fibrosarcoma) Whole Cell Lysate at 20 µg

      Secondary
      Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size: 88 kDa
      Observed band size: 130 kDa
      why is the actual band size different from the predicted?


      Exposure time: 20 minutes


      Integrin beta 1 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
    • Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)

      ab30394 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 ab30394 at 10μg/ml concentration and ab202272 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 Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labeled in blue with DAPI.

    • Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (ab30394)

      Overlay histogram showing HAP1 wildtype (green line) and HAP1-ITGB1 knockout cells (red line) stained with ab30394.

      Live HAP1 wildtype and HAP1-ITGB1 knockout cells were incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab30394, 1µg/0.5x106 cells) for 30 min at 22°C. A mouse IgG1 isotype control antibody (ab170190) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-ITGB1 knockout - grey line). Unlabeled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    • Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)This image is courtesy of an anonymous abreview.

      ab30394 at 1/500 staining Integrin beta 1 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by Immunocytochemistry/ Immunofluorescence.

      Cells were fixed in paraformaldehyde. The secondary used was an Alexa Fluor® 488 Goat anti mouse (H + L), used at a 1/500 dilution. Actin filaments (Phalloidin-TRITC) and DNA (DAPI, blue) are also shown

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)

      ab30394 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

      The cells were fixed in 4% formaldehyde for 10 minutes at room temperature and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30394 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was a Goat Anti-Mouse Alexa Fluor® 488 (IgG H&L) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    • Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (ab30394)

      Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab30394 (red line).

      The cells were fixed with 80% methanol (5 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 (ab30394, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) ab96879 at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)
      Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)This image is courtesy of an anonymous abreview.

      ab30394 at 10 µg/ml staining Integrin beta 1 in HT1080 (Human fibrosarcoma cell line) cells by Immunocytochemistry/ Immunofluorescence.

      Cells were fixed in paraformaldehyde. The secondary used was an Alexa Fluor® 555 Donkey anti mouse (H + L), used at a 1/500 dilution.

    References

    This product has been referenced in:

    • Jessen TN & Jessen JR VANGL2 protein stability is regulated by integrin av and the extracellular matrix. Exp Cell Res 374:128-139 (2019). Read more (PubMed: 30472097) »
    • Jacquemet G  et al. Filopodome Mapping Identifies p130Cas as a Mechanosensitive Regulator of Filopodia Stability. Curr Biol 29:202-216.e7 (2019). Read more (PubMed: 30639111) »
    See all 51 Publications for this product

    Publishing research using ab30394? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 27 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Integrin beta 1 antibody [12G10]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (OV90)
    Permeabilization
    No
    Specification
    OV90
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 24 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Integrin beta 1 antibody [12G10]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
    Sample
    Human Cell (Hela cell)
    Specification
    Hela cell
    Permeabilization
    Yes - 0.5% TritonX/PBS
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 19 2013

    Question

    Hello, I wanted to ask is this antibody recognizing activated b1 integrins?

    Read More
    Answer

    Yes, it does. Anti-Integrin beta 1 antibody ab30394 [12G10] is conformation-specific, and it recognizes the subset of beta1 integrins with high affinity for ligand (i.e. activated). In fact, it is the best antibody for this purpose because it binds close to the ligand-binding site, and the expression of its epitope is regulated by divalent cations in the same way that ligand binding is regulated.

    For more information about the production and characterization of the antibody, please consult the following reference.

    Identification of a novel anti-integrin monoclonal antibody that recognises a ligand-induced binding site epitope on the beta 1 subunit.
    FEBS Lett. 1995 Apr 17;363(1-2):118-22. PMID:7537221

    http://www.ncbi.nlm.nih.gov/pubmed/7537221?dopt=Abstract

    Read More

    Question

    Objective: I am interested in one of your R-PE conjugation kit for FACS antibodies.

    Previously, we ordered Integrin B1 antibody ((12G10)(ab30394)) for FACS analysis.

    Because your antibody is a purified one, I plan to conjugate R-PE dye to the antibodies and I found your R-PE conjugation kit (ab 102918).

    I want to know whether I can use your R-PE conjugation kit-produced labeled antibodies directly to the sample
    or I should use extra antibody purification step (such as column purification) before FACS analysis.

    Please let me know ASAP.

    Read More
    Answer

    Thank you for contacting us.

    We have the same integrin beta 1 antibody [12G10] that is FITC conjugated as ab150002, just so you know, but we do not have an R-PE conjugated version.

    But if you would like to conjugate the ab30394 to R-PE using our EasyLink kit ab102918, that would definitely work. Once the antibody is conjugated using the kit, it is ready to use and no further purification is necessary.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    I would like to follow up on the replacement antibody discussed below.

    Can we request a replacement?

    Regards,

    Read More
    Answer

    Thank you very much for your email.

    Looking through our records, it seems we already issued a credit to compensate for the ab30394 that wasn't working, as requested by email in May. The credit note number is ***, and it was probably applied back to the credit card used on the order. Could you confirm with your purchasing department whether this credit was received? They can contact our accounting department at mailto:us.credits@abcam.com or 888-772-2226 if they have any questions. This credit can be applied to the invoice of a new order placed with us, so it will essentially be a replacement antibody of your choice.

    I am very sorry for any confusion regarding this credit note, but please let me know if you have any questions or concerns, and I'll be happy to help you further.

    Read More

    Question

    AbID: 30394, Anti-Integrin beta 1 antibody [12G10] Rating: Inconclusive Image: Anti-Integrin beta 1 antibody [12G10] for Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) (Monkey) Sample: Species: Monkey Type: Tissue sections Specification: Eye Application: Application: Fixative: Paraformaldehyde Antigen retrieval step: None Permeabilization: Yes Blocking step: BSA as blocking agent for 1 hour at 23°C Other detail: Dilution: 1/100 Incubation time: 48 hours at 23°C Diluent: 1% BSA, 5% Donkey serum Secondary Antibody: Name: Non-Abcam Antibody was used: Alexa Fluor 555 donkey anti-mouse IgG Conjugation: Alexa Fluor® 555 Dilution: 1/200 Additional Data: Additional Notes: For picture: Blue=DAPI, Gray=Nomarski, Green=Integrin

    Read More
    Answer

    Thanks for submitting your review. I trust you have already been notified that your review has been "published" to our website.
    I just wanted to follow up with you since you gave an 'Inconclusive' rating for the review. You note using a previously untested application (IHC-P) and a previously untested specie (monkey) in your experiment. Can you please provide a bit more detail to explain the rating?
    Thanks very much! We really appreciate any additional detials you can provide.

    Read More

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Integrin beta 1 antibody [12G10]

     Inconclusive
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Monkey Tissue sections (Eye)
    Specification
    Eye
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    None
    Permeabilization
    Yes - Triton-X
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
    Read More

    Dr. Murray Johnstone

    Verified customer

    Submitted Aug 27 2012

    Question

    I want an antibody that recognizes total integrin B1. I know 12g10 recognizes the active conformation. Which will recognize both active and inactive. Thank you

    Read More
    Answer

    Thank you for contacting us.

    Several of the monoclonals we carry, mouse or rabbit, probably recognize total integrin beta 1, and their affinity is unlikely to be depend on whether an active conformation has been induced by ligand. However, they have not, to our knowledge, been characterized as thoroughly as the 12g10 clone was in this regard. FEBS Lett. 1995 Apr 17;363(1-2):118-22. PMID: 7537221.

    Of the other clones, the rabbit monoclonal ab52971 has been published several times and we have received many positive reviews from customers, Including some western blot reviews that suggest it is specific for an epitope common to several isoforms.

    To be assured that you are detecting total integrin beta 1, I recommend a rabbit polyclonal raised against a larger fragment, as it will detect multiple epitopes that are independent of binding of ligand to the integrin. For example, ab118206:

    Click here (or use the following: https://www.abcam.com/index.html?datasheet=118206).

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    Wl for the following product code. AB23750 AB11575 AB292 AB6586 AB30394 AB96879 AB96885. If the products does not requires MSDS, please certify the products are non-hazardous products for which no MSDS is provided or required. Thank you.

    Read More
    Answer

    r/>
    I can confirm the Safety Data Sheets can be found on the online datasheets. Near the top right hand side of the datasheet, there are two 'buttons', one for Datasheet PDF and the one underneath says SDS. Click on the SDS button to link to the document.

    I hope this will be helpful. If you have any difficulties finding these, or if you have any further questions, please do not hesitate to contact me.

    Read More

    Question

    We would be using the antibody for immunofluoresence and western blot
    preferably.

    Read More
    Answer

    Thank you for contacting us.

    The order has been placed for ab30394 with order number 1094397.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    1-10 of 27 Abreviews or Q&A

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