Anti-VE Cadherin抗体- Intercellular Junction Marker (ab33168)

概述

  • 产品名称

    Anti-VE Cadherin抗体- Intercellular Junction Marker
    参阅全部 VE Cadherin 一抗
  • 描述

    兔多克隆抗体to VE Cadherin - Intercellular Junction Marker
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC/IF, WB, IP, In-Cell ELISA, Flow Cytmore details
  • 种属反应性

    与反应: Mouse, Chicken, Human
    预测可用于: Cow, Pig
  • 免疫原

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

  • 阳性对照

    • WB: HUVEC Cell Lysate. ICC/IF: HUVEC cells. IHC-P: Mouse heart tissue. Flow Cyt: REH B cells. IP: HUVEC whole cell lysate.
  • 常规说明

      

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能

    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.
  • 组织特异性

    Endothelial tissues and brain.
  • 序列相似性

    Contains 5 cadherin domains.
  • 翻译后修饰

    Phosphorylated on tyrosine residues by KDR/VEGFR-2. Dephosphorylated by PTPRB.
  • 细胞定位

    Cell junction. Cell membrane. Found at cell-cell boundaries and probably at cell-matrix boundaries.
  • Information by UniProt
  • 数据库链接

  • 别名

    • 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

图片

  • Anti-VE Cadherin antibody - Intercellular Junction Marker (ab33168) at 1 µg/ml + HUVEC Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 87 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab33168 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

     

    The band we observe at 115 kDa is believed to be the glycosylated form of the protein.

  • Anti-VE Cadherin antibody - Intercellular Junction Marker (ab33168) at 1 µg/ml + HUVEC Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 87 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?
    Additional bands at: 55 kDa (possible non-specific binding)


    Exposure time: 1 minute


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab33168 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

     

    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

    This image was generated using confluent cells.

  • 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 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 stained HUVEC cells. The cells were incubated with the antibody ab33168 at 1/150 (Green). The cells were also stained with Rhodamine phalloidin (Red).

  • Anti-VE Cadherin antibody - Intercellular Junction Marker (ab33168) at 1 µg/ml + HUVEC Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 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 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

文献

This product has been referenced in:

  • Vaickus M  et al. Mild Traumatic Brain Injury in Mice Beneficially Alters Lung NK1R and Structural Protein Expression to Enhance Survival after Pseudomonas aeruginosa Infection. Am J Pathol 189:295-307 (2019). Read more (PubMed: 30472211) »
  • Li W & Zhou Y LRIG1 acts as a critical regulator of melanoma cell invasion, migration, and vasculogenic mimicry upon hypoxia by regulating EGFR/ERK-triggered epithelial-mesenchymal transition. Biosci Rep 39:N/A (2019). Read more (PubMed: 30487162) »
See all 115 Publications for this product

客户评价及客户问答

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1-10 of 37 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (liver, endothelial)
Permeabilization
No
Specification
liver, endothelial
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 23°C
Fixative
Paraformaldehyde

Miss. Kara Shumansky

Verified customer

提交于 Mar 11 2019

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

提交于 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

提交于 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

提交于 Sep 26 2017

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

提交于 Aug 02 2017

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

提交于 Aug 02 2017

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

提交于 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

提交于 May 01 2017

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

提交于 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

提交于 Mar 28 2017

1-10 of 37 Abreviews

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