MA5-29141

antibody from Invitrogen Antibodies

Targeting: CDH5 7B4, CD144

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-29141

Provider

Invitrogen Antibodies

Product name

VE-cadherin Recombinant Rabbit Monoclonal Antibody (48)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

This product is preservative free. It is recommended to add sodium azide to avoid contamination (final concentration 0.05%-0.1%).

Antibody clone number

48

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-VE-cadherin Recombinant Rabbit Monoclonal Antibody (48) (Product # MA5-29141) and ~65, 80,100,130 kDa band corresponding to Cadherin-5 was observed across high expressing cell lines (HUVEC and BeWo) tested, and not in low expressing cell lines (MCF7, OVCAR3, HeLa and Hep G2). Whole extracts (Scraped) (30 µg lysate) of HUVEC (Lane 1), MCF7 (Lane 2), OVCAR3 (Lane 3), HeLa (Lane 4), Hep G2 (Lane 5) and BeWo (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence staining of VE-cadherin in HUVEC cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with VE-cadherin Recombinant Rabbit Monoclonal Antibody (48) (Product # MA5-29141, 1:60) at 4°C overnight. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-rabbit IgG secondary antibody (green) and counterstained with DAPI (blue).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Cadherin-5 was performed using 90% confluent log phase HUVEC (Positive) and Hep G2 (Negative) cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with VE-cadherin Recombinant Rabbit Monoclonal Antibody (48) (Product # MA5-29141) at 1:50 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cell junction and plasma membrane localization. Panel e represents Hep G2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of Human CD144 expression on HUVEC cells. Cells were stained with VE-cadherin Recombinant Rabbit Monoclonal Antibody (48) (Product # MA5-29141), then a FITC-conjugated Secondary antibody. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells.