Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Integrin alpha 5 antibody [EPR7854] (ab150361)

Overview

  • Product name

    Anti-Integrin alpha 5 antibody [EPR7854]
    See all Integrin alpha 5 primary antibodies
  • Description

    Rabbit monoclonal [EPR7854] to Integrin alpha 5
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Integrin alpha 5 aa 1000-1100 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Human bladder, HT-1080, HeLa, and U937 cell lysates. IHC-P: Human kidney tissue. ICC/IF: U937, MCF7 and wild-type HAP1 cells. Flow Cyt: HeLa cells.
  • General notes

     

    A trial size is available to purchase for this antibody.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab150361 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 1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 115 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.
IP 1/10 - 1/100.
Flow Cyt 1/100 - 1/1000.

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

Target

  • 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
  • Database links

  • Alternative names

    • CD49 antigen-like family member E antibody
    • CD49e antibody
    • Fibronectin receptor subunit alpha antibody
    • Fibronectin receptor, alpha subunit antibody
    • FNRA antibody
    • Integrin alpha 5 (fibronectin receptor alpha) antibody
    • Integrin alpha-5 antibody
    • Integrin alpha-5 light chain antibody
    • Integrin alpha-F antibody
    • Integrin, alpha 5 (fibronectin receptor, alpha polypeptide) antibody
    • ITA5_HUMAN antibody
    • Itga5 antibody
    • Very late activation protein 5, alpha subunit antibody
    • VLA-5 antibody
    • VLA5 antibody
    • VLA5A antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Integrin alpha 5 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jeg3 cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab150361 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab150361 was shown to specifically recognize Integrin alpha 5 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when Integrin alpha 5 knockout samples were used. Wild-type and Integrin alpha 5 knockout samples were subjected to SDS-PAGE. ab150361 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1hr at room temperature before imaging.

  • Immunohistochemical analysis of paraffin embedded Normal Human Tonsil tissue using ab150361 showing +ve staining.

  • 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.

  • Overlay histogram showing HeLa cells stained with 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.

  • 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U937 (Human histiocytic lymphoma) cells labeling Integrin alpha 5 with 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).

  • Immunohistochemical analysis of paraffin embedded Human Brain vessels tissue using ab150361 showing +ve staining.

  • All lanes : Anti-Integrin alpha 5 antibody [EPR7854] (ab150361) at 1/2000 dilution

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Rat heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 115 kDa


    Exposure time: 3 minutes


    Observed band size : 19, 150 kDa

    Exposure time : 3 minutes

    Blocking/Diluting buffer 5% NFDM/TBST

  • All lanes : Anti-Integrin alpha 5 antibody [EPR7854] (ab150361) at 1/2000 dilution

    Lane 1 : Human bladder tissue lysate
    Lane 2 : HT-1080 (human fibrosarcoma) whole cell lysates
    Lane 3 : HeLa (human cervix adenocarcinoma) whole cell lysates
    Lane 4 : U937 (human monocyte histiocytic lymphoma) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 115 kDa



    Observed band size : 19, 150 kDa

    Exposure time : Lane 1-2: 1 minutes, Lane 3-4: 3 minutes

    Blocking/Diluting buffer 5% NFDM/TBST 

  • Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Integrin alpha 5 with ab150361 antibody at a dilution of 1/100.
  • Immunohistochemical analysis of paraffin embedded Human Vessels tissue using ab150361 showing +ve staining.

  • Immunohistochemical analysis of paraffin embedded Human Skeletal muscle vessel tissue using ab150361 showing +ve staining.

  • Immunohistochemical analysis of paraffin embedded Normal Human Uterus tissue using ab150361 showing +ve staining.

References

This product has been referenced in:

See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
20 µg
Specification
MEF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Mar 18 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Skin)
Antigen retrieval step
Other
Permeabilization
No
Specification
Skin
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 14 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (HUES7 ES cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
HUES7 ES cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Apr 14 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (undifferentiated Human ES cells on MEFs)
Permeabilization
Yes - see blocking buffer
Specification
undifferentiated Human ES cells on MEFs
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 23 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citrate
Permeabilization
No
Specification
Skin
Blocking step
Casein as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 23 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEPG2)
Specification
HEPG2
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 18 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
20 µg
Specification
HEPG2
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Nov 13 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 22 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Sample
Mouse Cell (mouse embryonic stem cells)
Specification
mouse embryonic stem cells
Permeabilization
Yes - 0.1% triton x
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 20 2013

Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Specification
mouse embryonic stem cell
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Nov 28 2013

1-10 of 12 Abreviews or Q&A

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