33-3900

antibody from Invitrogen Antibodies

Targeting: CDH2 CD325, CDHN, NCAD

Provider product page for 33-3900

Western blot

Antibody data

Antibody Data
Antigen structure
References [221]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [2]
Immunohistochemistry [1]
Flow cytometry [1]
Other assay [15]

Submit

Product number: 33-3900 - Provider product page
Provider: Invitrogen Antibodies
Product name: N-cadherin Monoclonal Antibody (3B9)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: This antibody is specific for N-cadherin and does not cross-react with other cadherin family members including P- and E-cadherins.
Antibody clone number: 3B9
Concentration: 0.5 mg/mL

CDH2 protein structure - 33-3900 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of N-Cadherin was performed by loading 20 µg of Rat Brain (lane1), Mouse Brain (lane2) and C2C12 (lane3) lysates using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800). Proteins were transferred to a PVDF membrane and blocked with 5 % skim milk for 1 hour at room temperature. N-Cadherin was detected at ~130 kDa using N-Cadherin Mouse Monoclonal Antibody (Product # 33-3900) at 1 µg - 3 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of total Cadherin and N-Cadherin was performed by loading 2 µL SeeBlue® Plus2 Prestained Protein Ladder (Product # LC5925), 50 µg of MDCK cell lysates and 10 µg mouse heart lysate per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 1% BSA/TBST for at least 1 hour at room temperature. Total cadherin was detected using a rabbit antibody (Product # 71-7100) and N-Cadherin was detected using a mouse antibody (Product # 33-3900), both at a concentration of 1 µg/mL in blocking buffer overnight at 4°C on a rocking platform. The blot was then incubated with goat anti-rabbit IgG-Alexa Fluor 790 secondary antibody (Product # A11369) and goat anti-mouse IgG-Alexa Fluor 680 secondary antibody (Product # A-21058) at a dilution of 1:10,000 for at least 1 hour. Fluorescent detection was performed using the Odyssey® CLx imaging system (Li-cor Biosciences). Image is generated by Joell Solan in Paul Lampe Lab at Fred Hutchinson Cancer Research Center.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of N-Cadherin was done on 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with N-Cadherin Mouse Monoclonal Antibody (Product # 33-3900) at 1 µg - 2 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938)and cytoskeletal F-actin (red) staining using Rhodamine Phalloidin (Product # R415, 1:300) panel c. Panel d is a merged image showing cell junctional localization of N-Cadherin. Panel e shows no primary antibody. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of N-Cadherin was done on 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with N-Cadherin Mouse Monoclonal Antibody (Product # 33-3900) at 1 µg - 2 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c is a merged image showing cell junctional localization of N-Cadherin. Panel d shows no primary antibody. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Re-morphogenesis of ARPE-19 cells treated with ITS. (A) The photomicrographs show ARPE-19 cells cultured for 4 weeks with ITS and stained for actin with Texas Red-X phalloidin. Note the tight intercellular contact, epithelial-like cellular shape, and predominantly peripheral cortical actin staining. Confocal immunofluorescence images of the same culture conditions showing beta-catenin (B) and N-cadherin (C) at the lateral membrane, CD147 (D) mostly in the apical domain and ezrin (E) in the apical domain. (F) The immunofluorescence image shows immunostaining of the alpha 1 subunit of Na + , K + -ATPase at the lateral membrane and more precisely, as observed in the XZ image, at the cell-cell contacts. (G) The beta 1 subunit is observed at the cell border and at the apical domain; the XZ image confirms both basolateral and apical staining. (H) The beta 2 subunit is observed mainly at the apical domain; the XZ image confirms apical staining. Scale bar: 10 mum. (I) Quantitative analysis of the transepithelial resistance (TER) of ARPE-19 cells during re-morphogenesis is depicted. Note a stable TER of ~80 Omega cm 2 between the second and fourth week in culture.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Rai14 is an apical and basal ES protein in the rat testis. Dual-labeled immunofluorescence analysis was used to assess co-localization of Rai14 (red) with constituent protein laminin-gamma3 chain (green), actin regulatory protein Arp3 (green), actin binding protein drebrin E (green) and actin filament cross-linking protein palladin (green) at the apical ES in stage VII tubules when these proteins were all highly expressed. It was found that Rai14 indeed partially co-localized with each of these apical ES proteins (see ""orange yellow"" in merged images) as shown in ( A ). Rai14 (red) also partially co-localized with basal ES/TJ protein ZO-1 (see ""yellow"" arrowheads), and to a lesser extent with N-cadherin (see ""white"" arrowheads) in stage IV-V tubules when these proteins were highly expressed as shown in ( B ). Bar = 10 um in ( A ) or ( B ), which applies all other images in ( A ) and ( B ).