Recombinant Anti-Integrin alpha 5 antibody [EPR7854] - Low endotoxin, Azide free (ab221606)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7854] to Integrin alpha 5 - Low endotoxin, Azide free
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Integrin alpha 5 antibody [EPR7854] - Low endotoxin, Azide free
See all Integrin alpha 5 primary antibodies -
Description
Rabbit monoclonal [EPR7854] to Integrin alpha 5 - Low endotoxin, Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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 (intra): HeLa cells. IP: HeLa cells.
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General notes
ab221606 is the carrier-free version of ab150361.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7854 -
Isotype
IgG -
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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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
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. -
Sequence similarities
Belongs to the integrin alpha chain family.
Contains 7 FG-GAP repeats. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 3678 Human
- Entrez Gene: 16402 Mouse
- Entrez Gene: 315346 Rat
- Omim: 135620 Human
- SwissProt: P08648 Human
- SwissProt: P11688 Mouse
- Unigene: 505654 Human
- Unigene: 16234 Mouse
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Alternative names
- CD49 antigen-like family member E antibody
- CD49e antibody
- Fibronectin receptor subunit alpha antibody
see all
Images
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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).
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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) -
Intracellular 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)
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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).
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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).
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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).
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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.
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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.
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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.
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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.
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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.
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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.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (3)
ab221606 has been referenced in 3 publications.
- Lopes-Ferreira M et al. Molecular Characterization and Functional Analysis of the Nattectin-like Toxin from the Venomous Fish Thalassophryne maculosa. Toxins (Basel) 14:N/A (2021). PubMed: 35050979
- Michalski D et al. Increased Immunosignals of Collagen IV and Fibronectin Indicate Ischemic Consequences for the Neurovascular Matrix Adhesion Zone in Various Animal Models and Human Stroke Tissue. Front Physiol 11:575598 (2020). PubMed: 33192578
- McMillan KJ et al. Atypical parkinsonism-associated retromer mutant alters endosomal sorting of specific cargo proteins. J Cell Biol 214:389-99 (2016). PubMed: 27528657