Why does the staining protocol with this Cadherin antibody use buffers containing Ca2+ and Mg2+?

Detects human VE-Cadherin in Western blots. In Western blots, 25% cross-reactivity with recombinant mouse VE‑Cadherin andno cross‑reactivity with recombinant human (rh) Cadherin-17 or rhP-Cadherin is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2B

Scientific Data Images for Human VE-Cadherin Antibody

Detection of Human VE‑Cadherin by Western Blot.

Western blot shows lysate of HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human VE-Cadherin Monoclonal Antibody (Catalog # MAB9381) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for VE-Cadherin at approximately 125 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

VE‑Cadherin in HUVEC Cells.

VE-Cadherin was detected in immersion fixed HUVEC cells using Mouse Anti-Human VE-Cadherin Monoclonal Antibody (Catalog # MAB9381) at 0.5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of VE‑Cadherin in HUVEC Human Cells by Flow Cytometry.

HUVEC human umbilical vein endothelial cells were stained with Mouse Anti-Human VE-Cadherin Monoclonal Antibody (Catalog # MAB9381, filled histogram) or isotype control antibody (MAB004, open histogram) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). View our protocol for Staining Membrane-associated Proteins.

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Applications for Human VE-Cadherin Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/10⁶ cells
Sample: HUVEC human umbilical vein endothelial cells stained in buffer containing Ca²⁺ and Mg²⁺

Immunocytochemistry

0.5-25 µg/mL
Sample: Immersion fixed HUVEC human umbilical vein endothelial cells

Western Blot

1 µg/mL
Sample: HUVEC human umbilical vein endothelial cells

Reviewed Applications

Read 2 reviews rated 4.5 using MAB9381 in the following applications:

Immunocytochemistry/Immunofluorescence (2 Reviews)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: VE-Cadherin

Vascular endothelial (VE)-cadherin (VE-CAD), also called 7B4 and cadherin‑5 (CDH5), is a member of the cadherin family of cell adhesion molecules. Cadherins are calcium‑dependent transmembrane proteins which bind to one another in a homophilic manner. On their cytoplasmic side, they associate with the three catenins, alpha, beta, and gamma (plakoglobin). This association links the cadherin protein to the cytoskeleton. Without association with the catenins, the cadherins are non-adhesive. Cadherins play a role in development, specifically in tissue formation. They may also help to maintain tissue architecture in the adult. VE-cadherin has been shown to play important roles in vasculogenesis and angiogenesis. VE-cadherin is a classical cadherin molecule. Classical cadherins consist of a large extracellular domain which contains DXD and DXNDN repeats responsible for mediating calcium-dependent adhesion, a single-pass transmembrane domain, and a short carboxy-terminal cytoplasmic domain responsible for interacting with the catenins. Human VE-cadherin is a 784 amino acid (aa) residue protein with a 25 aa signal sequence and a 759 aa propeptide. The mature protein begins at amino acid 48 and has a 546 aa extracellular domain, a 27 aa transmembrane domain, and a 164 aa cytoplasmic domain. The human and mouse mature VE-cadherin proteins share approximately 74% homology.

References

Shimoyama, Y. et al. (1989) J. Cell Biol. 109:1787.
Bussemakers, M.J.G. et al. (1993) Mol. Biol. Reports 17:123.
Overduin, M. et al. (1995) Science 267:386.
Takeichi, M. (1991) Science 251:1451.
Nose, A. et al. (1987) EMBO J. 6:3655.
Carmeliet, P. et al. (1999) Cell 98:147.
Gory-Faure, S. et al. (1999) Development 126:2093.

Long Name

Vascular Endothelium Cadherin

Alternate Names

Cadherin-5, CD144, CDH5, VECadherin

Entrez Gene IDs

1003 (Human); 12562 (Mouse)

Gene Symbol

CDH5

UniProt

P33151

Additional VE-Cadherin Products

Product Documents for Human VE-Cadherin Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human VE-Cadherin Antibody

For research use only

Citations
Reviews

Protocols
FAQs

Calculators
Flow Cytometry

Human VE-Cadherin Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB9381

Key Product Details

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Species Reactivity

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Applications