Overview

  • Product name
    Anti-VE Cadherin antibody - Intercellular Junction Marker
    See all VE Cadherin primary antibodies
  • Description
    Rabbit polyclonal to VE Cadherin - Intercellular Junction Marker
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IP, In-Cell ELISA, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Chicken, Human
    Predicted to work with: Cow, Pig
  • Immunogen

    Synthetic peptide corresponding to Human VE Cadherin aa 750 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab27462)

  • Positive control
    • WB: HUVEC Cell Lysate. ICC/IF: HUVEC cells. IHC-P: Mouse heart tissue. Flow Cyt: REH B cells. IP: HUVEC whole cell lysate.
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab33168 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 0.1 - 1 µg/ml.

Abcam recommends using this product with confluent cells.

WB Use a concentration of 1 µg/ml. Detects a band of approximately 115 kDa (predicted molecular weight: 87 kDa).Can be blocked with VE Cadherin peptide (ab27462).

Abcam recommends using BSA blocking with this product.  Milk blocking will give a greatly reduced signal strength in WB.

IP Use at an assay dependent concentration.
In-Cell ELISA Use at an assay dependent concentration. PubMed: 22689949
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Cadherins are calcium dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. This cadherin may play a important role in endothelial cell biology through control of the cohesion and organization of the intercellular junctions. It associates with alpha-catenin forming a link to the cytoskeleton.
  • Tissue specificity
    Endothelial tissues and brain.
  • Sequence similarities
    Contains 5 cadherin domains.
  • Post-translational
    modifications
    Phosphorylated on tyrosine residues by KDR/VEGFR-2. Dephosphorylated by PTPRB.
  • Cellular localization
    Cell junction. Cell membrane. Found at cell-cell boundaries and probably at cell-matrix boundaries.
  • Information by UniProt
  • Database links
  • Alternative names
    • 7B 4 antibody
    • 7B4 antibody
    • 7B4 antigen antibody
    • CADH5_HUMAN antibody
    • Cadherin 5 antibody
    • Cadherin 5 type 2 antibody
    • Cadherin 5, type 2 (vascular endothelium) antibody
    • Cadherin 5, type 2, VE cadherin (vascular epithelium) antibody
    • cadherin, vascular endothelial antibody
    • cadherin, vascular endothelial, 1 antibody
    • Cadherin-5 antibody
    • Cadherin5 antibody
    • CD 144 antibody
    • CD144 antibody
    • CD144 antigen antibody
    • CDH 5 antibody
    • CDH5 antibody
    • CDH5 protein antibody
    • Endothelial specific cadherin antibody
    • FLJ17376 antibody
    • OTTHUMP00000174777 antibody
    • Vascular endothelial cadherin antibody
    • Vascular epithelium cadherin antibody
    • VE Cad antibody
    • VE-cadherin antibody
    • VEC antibody
    see all

Images

  • Anti-VE Cadherin antibody - Intercellular Junction Marker (ab33168) at 1 µg/ml + HUVEC Cell Lysate at 20 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 87 kDa
    Observed band size: 115 kDa
    why is the actual band size different from the predicted?



    The band we observe at 115 kDa is believed to be the glycosylated form of the protein.
  • ab33168 stained in HUVEC cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab33168 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

  • ab33168 at 1 µg/ml + HUVEC at 10 µg

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/1000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 87 kDa
    Observed band size: 115,117 kDa why is the actual band size different from the predicted?
    Additional bands at: 45 kDa (possible non-specific binding)


    Exposure time: 1 minute


    The observed band for Cadherin 5 has a higher molecular weight of 115kDa due to glycosylation of the protein. 

    The immunogen used to raise this antibody has 89% homology with Cadherin 18, 88kDa , which we believe is the additional observed band at 117kDa, again due to glycosylation of the protein. 

  • ab33168 used in Flow cytometry.
    Human REH B cells were fixed in paraformaldehyde and permeabilized using saponin. Primary antibody used undiluted (2µl in 100µl of cells in PBS) and incubated for 15 minutes at 4°C. The secondary antibody used was an undiluted, Alexa Fluor®488 conjugated goat anti-rabbit IgG.

    Rabbit IgG isotype control (white)

    See Abreview

  • ICC/IF image of VE-Cadherin staining on HUVEC cells using ab33168. The cells were incubated with the primary antibody (ab33168) and the secondary was FITC conjugated anti-rabbit used at 1:400. The cells were incubated with only the secondary antibody as a negative control.
  • ab33168 Immunoprecipitating VE Cadherin in human HUVEC whole cell lysate. 1000000 cells lysate was incubated with primary antibody (1/100 in 0.5% NP40, 150mM NaCl, 50mM Tris) and matrix (Dynabeads) for 2 hours at 4°C. For western blotting a HRP-conjugated mouse anti-VE Cadherin (1/3000) was used to confirm successful immunoprecipation.

    See Abreview

  • ICC/IF image of VE Cadherin stained HUVEC cells. The cells were incubated with the antibody ab33168 at 1/150 (Green). The cells were also stained with Rhodamine phalloidin (Red).

References

This product has been referenced in:
  • Soni D  et al. Deubiquitinase function of A20 maintains and repairs endothelial barrier after lung vascular injury. Cell Death Discov 4:60 (2018). Read more (PubMed: 29796309) »
  • Miscianinov V  et al. MicroRNA-148b Targets the TGF-ß Pathway to Regulate Angiogenesis and Endothelial-to-Mesenchymal Transition during Skin Wound Healing. Mol Ther N/A:N/A (2018). WB, ICC/IF . Read more (PubMed: 29843955) »
See all 89 Publications for this product

Customer reviews and Q&As

1-10 of 51 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Pig Cultured Cells (Endothelial cell)
Permeabilization
Yes - Methanol
Specification
Endothelial cell
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Oct 10 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Bos taurus Cell (Primary endothelial cells)
Permeabilization
Yes - 0.25% Triton X-100
Specification
Primary endothelial cells
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 26 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Embryo)
Permeabilization
No
Specification
Embryo
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 26 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human umbilical vein endothelial cell)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Loading amount
20 µg
Treatment
siRNA for 48h
Specification
human umbilical vein endothelial cell
Blocking step
Milk as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: ~25°C

Abcam user community

Verified customer

Submitted Aug 02 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (liver)
Permeabilization
Yes - 0.5% TritonX-100
Specification
liver
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Dr. shuang liang

Verified customer

Submitted Aug 02 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Hamster Tissue sections (Heart)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9.0
Permeabilization
No
Specification
Heart
Blocking step
BSA + Milk + Serum as blocking agent for 20 minute(s) · Concentration: 2.0% · Temperature: 27°C
Fixative
Formaldehyde

Dr. Marcelo Pelajo Machado

Verified customer

Submitted Jul 21 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Primary mouse lung endothelial cells)
Permeabilization
No
Specification
Primary mouse lung endothelial cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Fixative
Paraformaldehyde

Dr. Jim Hsiao

Verified customer

Submitted May 01 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human umbilical vein endothelial cell)
Permeabilization
Yes - 0.5% TritonX-100
Specification
human umbilical vein endothelial cell
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Dr. shuang liang

Verified customer

Submitted Mar 31 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (human umbilical vein endothelial cell)
Gel Running Conditions
Non-reduced Denaturing (10% gel)
Loading amount
20 µg
Treatment
siRNA for 48hrs
Specification
human umbilical vein endothelial cell
Blocking step
Milk as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: ~25°C

Dr. 振洋 余

Verified customer

Submitted Mar 28 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human umbilical vein endothelial cell)
Permeabilization
Yes - 0.5% Triton X-100 for 20min
Specification
human umbilical vein endothelial cell
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde

Dr. 振洋 余

Verified customer

Submitted Mar 24 2017

1-10 of 51 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up