Anti-Integrin beta 1 antibody [12G10] (ab30394)
![Western blot - Anti-Integrin beta 1 antibody [12G10] (ab30394)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-40377-anti-integrin-beta-1-antibody-12g10-western-blot.jpg "Western blot - Anti-Integrin beta 1 antibody [12G10] (ab30394)")
Key features and details
- Mouse monoclonal [12G10] to Integrin beta 1
- Suitable for: ICC/IF, Flow Cyt, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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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, 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.
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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. ICC: HeLa cells
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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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
Purified from tissue culture supernatant. -
Primary antibody notes
This antibody also enhances alpha 5 - beta 1 - fibronectin interactions. -
Clonality
Monoclonal -
Clone number
12G10 -
Myeloma
x63-Ag8.653 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 | (3) |
Use a concentration of 1 - 10 µg/ml.
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| Flow Cyt | (1) |
Use a concentration of 1 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
| WB | (2) |
Use a concentration of 5 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 88 kDa).
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| Notes |
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ICC/IF
Use a concentration of 1 - 10 µg/ml. |
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Flow Cyt
Use a concentration of 1 µg/ml. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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WB
Use a concentration of 5 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 88 kDa). |
Target
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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
modificationsThe 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
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Database links
- Entrez Gene: 3688 Human
- Omim: 135630 Human
- SwissProt: P05556 Human
- Unigene: 643813 Human
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Alternative names
- beta1 integrin antibody
- CD_antigen=CD29 antibody
- CD29 antibody
see all
Images
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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)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-261759-anti-integrin-beta-1-antibody-12g10-immunofluorescence.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-13-anti-integrin-beta-1-12g10-antibody-immunocytochemistry-hela-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)ab30394 staining Integrin beta 1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 overnight at 4°C with ab30394 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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![Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (ab30394)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-276817-anti-integrin-beta-1-antibody-12g10-flow-cytometry.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-211311-anti-integrin-beta-1-antibody-12g10-immunofluorescence.jpg)
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
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![Flow Cytometry - Anti-Integrin beta 1 antibody [12G10] (ab30394)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-40376-anti-integrin-beta-1-antibody-12g10-flow-cytometry.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Integrin beta 1 antibody [12G10] (ab30394)](https://www.abcam.com/ps/products/30/ab30394/Images/ab30394-40378-anti-integrin-beta-1-antibody-12g10-immunofluorescence.jpg)
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (82)
ab30394 has been referenced in 82 publications.
- Xie J et al. Energy expenditure during cell spreading influences the cellular response to matrix stiffness. Biomaterials 267:120494 (2021). PubMed: 33130323
- Wang Y et al. Positive effect of Astragaloside IV on neurite outgrowth via talin-dependent integrin signaling and microfilament force. J Cell Physiol 236:2156-2168 (2021). PubMed: 32853433
- Takeuchi M et al. Neutrophils interact with cholangiocytes to cause cholestatic changes in alcoholic hepatitis. Gut 70:342-356 (2021). PubMed: 33214166
- Gutjahr JC et al. CD44 engagement enhances acute myeloid leukemia cell adhesion to the bone marrow microenvironment by increasing VLA-4 avidity. Haematologica 106:2102-2113 (2021). PubMed: 32616529
- Tu WJ et al. Targeting novel LSD1-dependent ACE2 demethylation domains inhibits SARS-CoV-2 replication. Cell Discov 7:37 (2021). PubMed: 34031383